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<article doi="10.1002/embj.201385902" abstract="Mitochondrial dysfunction has long been associated with Parkinson's disease (PD). Parkin and PINK1, two genes associated with familial PD, have been implicated in the degradation of depolarized mitochondria via autophagy (mitophagy). Here, we describe the involvement of parkin and PINK1 in a vesicular pathway regulating mitochondrial quality control. This pathway is distinct from canonical mitophagy and is triggered by the generation of oxidative stress from within mitochondria. Wild-type but not PD-linked mutant parkin supports the biogenesis of a population of mitochondria-derived vesicles (MDVs), which bud off mitochondria and contain a specific repertoire of cargo proteins. These MDVs require PINK1 expression and ultimately target to lysosomes for degradation. We hypothesize that loss of this parkin- and PINK1-dependent trafficking mechanism impairs the ability of mitochondria to selectively degrade oxidized and damaged proteins leading, over time, to the mitochondrial dysfunction noted in PD."><fig id="1125"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1125"/><sd-panel panel_id="2247"><div>B <sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells expressing <sd-tag id="sdTag2" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (green), <sd-tag id="sdTag5" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OCT" ext_ids="P00480" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OTC" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">OCT</sd-tag>-<sd-tag id="sdTag4" source="sdapi" category="entity" entity_type="protein" role="reporter" text="DsRed2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DsRed2</sd-tag> (red) and <sd-tag id="sdTag6" source="sdapi" category="entity" entity_type="protein" role="component" text="CFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">CFP</sd-tag>-<sd-tag id="sdTag468" source="sdapi" category="entity" entity_type="protein" role="component" text="Drp1" ext_ids="O00429" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Drp1</sd-tag>K38E were left untreated (untreated) or treated for <sd-tag id="sdTag81" source="sdapi" category="time" entity_type="" role="" text="2 h" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">2 h</sd-tag> with 10 µM <sd-tag id="sdTag8" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag> (<sd-tag id="sdTag9" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag>), then fixed and stained for <sd-tag id="sdTag10" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue). Scale bars, 30 µm.</div><graphic href="https://api.sourcedata.io/file.php?panel_id=2247"/></sd-panel><sd-panel panel_id="2248">C Cells prepared as in (B) were treated with 50 µM <sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> for 2 h. Arrows indicate <sd-tag id="sdTag12" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OCT" ext_ids="P00480" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OTC" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">OCT</sd-tag>-<sd-tag id="sdTag13" source="sdapi" category="entity" entity_type="protein" role="reporter" text="DsRed2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DsRed2</sd-tag>-positive/<sd-tag id="sdTag14" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>-negative <sd-tag id="sdTag26" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> that colocalize with <sd-tag id="sdTag15" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag16" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>, while circles indicate <sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> that are <sd-tag id="sdTag17" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>-negative. Arrowheads indicate <sd-tag id="sdTag385" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="nascent vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">nascent <sd-pretag id="sdPretag14237323342560" parent-tag-id="392" class="component subcellular">vesicles</sd-pretag></sd-tag>, adjacent to <sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>, exhibiting cargo selectivity and <sd-tag id="sdTag18" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> recruitment. Open arrowheads indicate <sd-tag id="sdTag19" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>-negative <sd-tag id="sdTag28" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> containing the reciprocal cargo (<sd-tag id="sdTag20" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>-positive/<sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OCT" ext_ids="P00480" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OTC" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">OCT</sd-tag>-<sd-tag id="sdTag22" source="sdapi" category="entity" entity_type="protein" role="reporter" text="DsRed2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DsRed2</sd-tag>-negative). Cell boundaries are delineated in the <sd-tag id="sdTag23" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag24" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> single-channel image. Scale bars, 30 µm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2248"/></sd-panel></fig><fig id="1126"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1126"/><sd-panel panel_id="2249">A Quantification of the number of <sd-tag id="sdTag30" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OCT" ext_ids="P00480" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OTC" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">OCT</sd-tag>μ<sd-tag id="sdTag31" source="sdapi" category="entity" entity_type="protein" role="reporter" text="DsRed2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DsRed2</sd-tag>μpositive/<sd-tag id="sdTag32" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>μnegative <sd-tag id="sdTag413" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag> in cells expressing <sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>, <sd-tag id="sdTag34" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ<sd-pretag id="sdPretag14528496738510" parent-tag-id="49" class="intervention gene">parkin</sd-pretag>WT, <sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ<sd-tag id="sdTag49" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag>R42P, <sd-tag id="sdTag38" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ<sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag>K211N, or <sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ<sd-tag id="sdTag51" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag>C431F, treated with or without <sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A) as in (C); both the total number (white bars) and the number colocalizing with <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ<sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Bars represent the mean ± s.e.m. Pμvalues are given first for <sd-tag id="sdTag45" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ/<sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>μ<sd-tag id="sdTag47" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>μpositive <sd-tag id="sdTag415" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag>, then for total <sd-tag id="sdTag416" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 49-68 cells in 2-3 experiments); ns, not significant; **P < 0.01; ***P < 0.001).<graphic href="https://api.sourcedata.io/file.php?panel_id=2249"/></sd-panel><sd-panel panel_id="2251">C <sd-tag id="sdTag56" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells expressing various <sd-tag id="sdTag57" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> mutant contructs (green), p<sd-tag id="sdTag55" source="sdapi" category="entity" entity_type="protein" role="assayed" text="OCT" ext_ids="P00480" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="OTC" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">OCT</sd-tag>-<sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="protein" role="reporter" text="DsRed2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">DsRed2</sd-tag> (red) and <sd-tag id="sdTag60" source="sdapi" category="entity" entity_type="protein" role="component" text="CFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">CFP</sd-tag>-<sd-tag id="sdTag66" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag>K38E were treated with 50 µM <sd-tag id="sdTag61" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> for <sd-tag id="sdTag387" source="sdapi" category="time" entity_type="" role="" text="2 h" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">2 h</sd-tag>, then fixed and <sd-tag id="sdTag386" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> against <sd-tag id="sdTag62" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>. Arrows indicate matrix-positive/<sd-tag id="sdTag63" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>-negative structures colocalizing with <sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag65" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>. Scale bars, 30 µm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2251"/></sd-panel><sd-panel panel_id="2252">D <sd-tag id="sdTag82" source="sdapi" category="assay" entity_type="" role="" text="Immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Immunoblot</sd-tag> of whole-cell lysates of <sd-tag id="sdTag67" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells expressing various <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="protein" role="component" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> constructs, treated with 50 µM <sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A, +) or <sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> (-) for <sd-tag id="sdTag389" source="sdapi" category="time" entity_type="" role="" text="2 h" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">2 h</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=2252"/></sd-panel></fig><fig id="1127"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1127"/><sd-panel panel_id="2253">A Representative <sd-tag id="sdTag86" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag> of whole‐cell lysates from <sd-tag id="sdTag120" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag87" source="sdapi" category="entity" entity_type="protein" role="component" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> and <sd-tag id="sdTag119" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag88" source="sdapi" category="entity" entity_type="protein" role="component" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag89" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> cells transfected with non‐targeting siRNA or siRNA targeting <sd-tag id="sdTag390" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag> (siDrp1).<graphic href="https://api.sourcedata.io/file.php?panel_id=2253"/></sd-panel><sd-panel panel_id="2254">B <sd-tag id="sdTag121" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="gene" role="component" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> cells transfected with siRNA targeting <sd-tag id="sdTag95" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag> were treated with <sd-tag id="sdTag96" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> (upper panels) or 25 μM <sd-tag id="sdTag97" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A, lower panels) for <sd-tag id="sdTag428" source="sdapi" category="time" entity_type="" role="" text="90 min" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">90 min</sd-tag>, then fixed and <sd-tag id="sdTag103" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> against <sd-pretag id="sdPretag14377433459440" parent-tag-id="456" class="component protein">PDH</sd-pretag> (red) and <sd-tag id="sdTag98" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue). <sd-tag id="sdTag105" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3 bp" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/"><sd-pretag id="sdPretag14236724445360" parent-tag-id="344" class="component protein">PDH</sd-pretag><sd-pretag id="sdPretag14376517840540" parent-tag-id="443">E2/E3</sd-pretag> bp</sd-tag>−positive/<sd-tag id="sdTag99" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative <sd-tag id="sdTag100" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> colocalizing with <sd-tag id="sdTag101" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag102" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (arrows) or not (circles) are indicated. Scale bars, 20 μm (first panels on left) and 2 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2254"/></sd-panel><sd-panel panel_id="2255">C Quantification of PDH−positive/<sd-tag id="sdTag107" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative <sd-tag id="sdTag417" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag> in <sd-tag id="sdTag118" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag108" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> and <sd-tag id="sdTag123" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag109" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag110" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> cells transfected with the indicated siRNA, treated with <sd-tag id="sdTag111" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> or <sd-tag id="sdTag112" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A) as in (B); both the total number (white bars) and the number colocalizing with <sd-tag id="sdTag113" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag114" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Bars represent the mean ± s.e.m. P−values are given first for <sd-tag id="sdTag115" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−/<sd-tag id="sdTag116" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag117" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>−positive <sd-tag id="sdTag418" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag>, then for total <sd-tag id="sdTag419" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 48-85 cells in 2-3 experiments); ns, not significant; ***P < 0.001).<graphic href="https://api.sourcedata.io/file.php?panel_id=2255"/></sd-panel><sd-panel panel_id="2256">D 5 μm‐long profile of the <sd-tag id="sdTag125" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>‐positive <sd-tag id="sdTag420" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> and adjacent <sd-tag id="sdTag127" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondrial tubule" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/"><sd-pretag id="sdPretag14376512290920" parent-tag-id="437" class="component subcellular">mitochondrial</sd-pretag> tubule</sd-tag> depicted in the <sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag>‐treated cell from (B) (left). Increasing position values on the x‐axis of the fluorescence intensity plot (right) correspond to moving from the top to the bottom of the profile. *<sd-tag id="sdTag130" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDV" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDV</sd-tag> indicated by an arrow in (B).<graphic href="https://api.sourcedata.io/file.php?panel_id=2256"/></sd-panel><sd-panel panel_id="2257">E <sd-tag id="sdTag394" source="sdapi" category="assay" entity_type="" role="" text="Quantification of the distance" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Quantification of the distance</sd-tag> between the centres of <sd-tag id="sdTag132" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag133" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag>−negative (black bar) or −positive (gray bar) <sd-tag id="sdTag393" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag> and the edge of the nearest <sd-tag id="sdTag469" source="sdapi" category="entity" entity_type="subcellular" role="component" text="mitochondrial" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondrial</sd-tag> tubule for the <sd-tag id="sdTag392" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag> quantified in <sd-tag id="sdTag135" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag>−treated <sd-tag id="sdTag136" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag137" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag138" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> cells transfected with si<sd-tag id="sdTag131" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag> in (C). Error bars represent the mean ± s.e.m.; *P < 0.05 (obtained by Student's t−test).<graphic href="https://api.sourcedata.io/file.php?panel_id=2257"/></sd-panel><sd-panel panel_id="2258">F <sd-tag id="sdTag142" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag143" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag144" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> cells transfected with siRNA targeting <sd-tag id="sdTag145" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag> (siDrp1) were treated with 25 μM <sd-tag id="sdTag146" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> for 90 min prior to fixation. Samples were <sd-tag id="sdTag147" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> against <sd-tag id="sdTag148" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue) and the indicated <sd-pretag id="sdPretag14376512428530" parent-tag-id="437">mitochondrial</sd-pretag> marker (red). <sd-tag id="sdTag149" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>‐negative <sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> containing the specified cargo and colocalizing with <sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (arrows) or not (circles) are indicated. Arrowheads show lack of colocalization between <sd-tag id="sdTag153" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> and the indicated <sd-pretag id="sdPretag14376512296620" parent-tag-id="437">mitochondrial</sd-pretag> marker. Scale bars, 20 μm (first two columns of panels) and 2 μm (remaining columns of panels).<graphic href="https://api.sourcedata.io/file.php?panel_id=2258"/></sd-panel><sd-panel panel_id="2259">G Quantification of <sd-tag id="sdTag160" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative structures that stained positively for the indicated cargo in <sd-tag id="sdTag161" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag162" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> (<sd-tag id="sdTag163" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>) and <sd-tag id="sdTag164" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag165" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag166" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> (<sd-tag id="sdTag167" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag168" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag>) cells, transfected with siRNA targeting <sd-tag id="sdTag169" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag>, treated with <sd-tag id="sdTag170" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A); both the total number (white bars) and the number colocalizing with <sd-tag id="sdTag171" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag172" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Error bars represent the mean ± s.e.m. P−values are given first for <sd-tag id="sdTag173" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−/<sd-tag id="sdTag174" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag175" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>−positive <sd-tag id="sdTag395" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag>, then for total <sd-tag id="sdTag396" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 24-59 cells in two experiments; ns, not significant; ***P < 0.001).<graphic href="https://api.sourcedata.io/file.php?panel_id=2259"/></sd-panel></fig><fig id="1128"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1128"/><sd-panel panel_id="2260">A<sd-tag id="sdTag190" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells, transfected with siRNA targeting <sd-tag id="sdTag191" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag> and <sd-tag id="sdTag192" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag193" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (green), were treated with 25 μM <sd-tag id="sdTag194" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> for <sd-tag id="sdTag397" source="sdapi" category="time" entity_type="" role="" text="90 min" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">90 min</sd-tag> following a <sd-tag id="sdTag398" source="sdapi" category="time" entity_type="" role="" text="30‐min" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">30‐min</sd-tag> pretreatment with 50 nM <sd-tag id="sdTag203" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="bafilomycin A1" ext_ids="CHEBI:22689" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="bafilomycin A1" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">bafilomycin A1</sd-tag>, then fixed and <sd-tag id="sdTag195" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> against <sd-tag id="sdTag454" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/"><sd-pretag id="sdPretag14377433362450" parent-tag-id="456">PDH</sd-pretag> E2/E3</sd-tag> bp (red) and <sd-tag id="sdTag197" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue). <sd-tag id="sdTag455" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/"><sd-pretag id="sdPretag14377433048810" parent-tag-id="456">PDH</sd-pretag> E2/E3</sd-tag> bp‐positive/<sd-tag id="sdTag199" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>‐negative <sd-tag id="sdTag200" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> colocalizing with <sd-tag id="sdTag201" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag202" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (arrows) or not (circles) are indicated. Scale bar, 30 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2260"/></sd-panel><sd-panel panel_id="2261">B Quantification of <sd-tag id="sdTag211" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3bp" ext_ids="P10515///P09622" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLAT///DLD" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3bp</sd-tag>‐positive/<sd-tag id="sdTag205" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>‐negative <sd-tag id="sdTag400" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag> in <sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells treated with <sd-tag id="sdTag207" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> in the presence of the lysosomal inhibitors <sd-tag id="sdTag208" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="bafilomycin A1" ext_ids="CHEBI:22689" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="bafilomycin A1" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">bafilomycin A1</sd-tag> or <sd-tag id="sdTag215" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="pepstatin A" ext_ids="CHEBI:7989" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Pepstatin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">pepstatin A</sd-tag> and <sd-tag id="sdTag216" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="E‐64d" ext_ids="CHEBI:30270" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="E64" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">E‐64d</sd-tag>; both total number (white bars) and the number colocalizing with <sd-tag id="sdTag209" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag210" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Bars represent the mean ± s.e.m. P‐values are given for total <sd-tag id="sdTag401" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 19 to 120 cells in 2-3 experiments; **P < 0.01; ***P < 0.001).<graphic href="https://api.sourcedata.io/file.php?panel_id=2261"/></sd-panel><sd-panel panel_id="2262">C<sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells, transfected with <sd-tag id="sdTag218" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag219" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (green) and siRNA targeting <sd-tag id="sdTag220" source="sdapi" category="entity" entity_type="gene" role="component" text="Drp1" ext_ids="10059" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="DNM1L" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Drp1</sd-tag> and <sd-tag id="sdTag230" source="sdapi" category="entity" entity_type="gene" role="component" text="Atg5" ext_ids="9474" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Atg5</sd-tag>, were treated with 25 μM <sd-tag id="sdTag221" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> for 90 min, then fixed, <sd-tag id="sdTag222" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> for <sd-tag id="sdTag456" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH</sd-tag> (red) and <sd-tag id="sdTag224" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue), and counterstained for <sd-tag id="sdTag232" source="sdapi" category="assay" entity_type="" role="" text="Hoescht" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Hoescht</sd-tag> (gray). <sd-tag id="sdTag233" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3bp" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3bp</sd-tag> bp‐positive/<sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>‐negative <sd-tag id="sdTag227" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> colocalizing with <sd-tag id="sdTag228" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag229" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (arrows) or not (circles) are indicated. Scale bars, 20 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2262"/></sd-panel><sd-panel panel_id="2263">D Quantification of <sd-tag id="sdTag243" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3</sd-tag> bp‐positive/<sd-tag id="sdTag235" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>‐negative <sd-tag id="sdTag402" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag> from <sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells transfected with the indicated siRNA; both the total number (white bars) and the number colocalizing with <sd-tag id="sdTag237" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag238" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Error bars represent the mean ± s.e.m. P‐values are given for the total <sd-tag id="sdTag403" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 42-64 cells in 2-3 experiments); ns, not significant.<graphic href="https://api.sourcedata.io/file.php?panel_id=2263"/></sd-panel></fig><fig id="1129"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1129"/><sd-panel panel_id="2264">A Representative <sd-tag id="sdTag245" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag> of whole‐cell lysates from <sd-tag id="sdTag246" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells transfected with non‐targeting siRNA or siRNA targeting <sd-tag id="sdTag248" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PINK1" ext_ids="65018" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PINK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PINK1</sd-tag> (siPINK1), treated with 10 μM <sd-tag id="sdTag247" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag> for 6 h in order to stabilize the <sd-tag id="sdTag249" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PINK1" ext_ids="65018" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PINK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PINK1</sd-tag> full‐length band.<graphic href="https://api.sourcedata.io/file.php?panel_id=2264"/></sd-panel><sd-panel panel_id="2265">B <sd-tag id="sdTag253" source="sdapi" category="entity" entity_type="cell_line" role="component" text="HeLa" ext_ids="CVCL_0030" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HeLa" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">HeLa</sd-tag> cells transfected with <sd-tag id="sdTag254" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag255" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (green) and siRNA targeting <sd-tag id="sdTag256" source="sdapi" category="entity" entity_type="gene" role="intervention" text="PINK1" ext_ids="65018" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PINK1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">PINK1</sd-tag> (siPINK1) or non−targeting control (ctrl siRNA), were treated with 25 μM <sd-tag id="sdTag257" source="sdapi" category="entity" entity_type="molecule" role="component" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> for <sd-tag id="sdTag442" source="sdapi" category="time" entity_type="" role="" text="90 min" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">90 min</sd-tag>, then fixed and <sd-tag id="sdTag258" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> for <sd-tag id="sdTag459" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3</sd-tag> bp (red) and <sd-tag id="sdTag260" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue). <sd-tag id="sdTag458" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3</sd-tag> bp−positive/<sd-tag id="sdTag262" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative <sd-tag id="sdTag263" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> colocalizing with <sd-tag id="sdTag264" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag265" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (arrows) or not (circles) are indicated. Arrowheads indicate <sd-tag id="sdTag266" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>−positive <sd-tag id="sdTag267" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MDVs" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">MDVs</sd-tag> adjacent to <sd-tag id="sdTag268" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>, possibly budding. Scale bar, 20 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2265"/></sd-panel><sd-panel panel_id="2266">C Quantification of <sd-tag id="sdTag457" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDHE2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDHE2/E3</sd-tag> bp−positive/<sd-tag id="sdTag270" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative structures from (B); both the total number (white bars) and the number colocalizing with <sd-tag id="sdTag271" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag272" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Error bars represent the mean ± s.e.m. P−values are given first for <sd-tag id="sdTag273" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−/<sd-tag id="sdTag274" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag275" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>−positive <sd-tag id="sdTag404" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag>, then for total <sd-tag id="sdTag405" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 42-60 cells in three experiments); ns, not significant; ***P < 0.001).<graphic href="https://api.sourcedata.io/file.php?panel_id=2266"/></sd-panel></fig><fig id="1130"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1130"/><sd-panel panel_id="2267">A Quantification of <sd-tag id="sdTag409" source="sdapi" category="assay" entity_type="" role="" text="TMRM fluorescence" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">TMRM fluorescence</sd-tag> by <sd-tag id="sdTag407" source="sdapi" category="assay" entity_type="" role="" text="flow cytometry" ext_ids="BAO_0000005" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="flow cytometry" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">flow cytometry</sd-tag> in <sd-tag id="sdTag279" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag280" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag281" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> cells treated with 25 μM <sd-tag id="sdTag282" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A), 25 μM <sd-tag id="sdTag283" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> and 10 μM <sd-tag id="sdTag408" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="oligomycin" ext_ids="CHEBI:25675" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="oligomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">oligomycin</sd-tag> (anti A + oligo), and 20 μM <sd-tag id="sdTag284" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag> (<sd-tag id="sdTag285" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag>) for the indicated time, represented as a fraction of fluorescence intensity compared to <sd-tag id="sdTag286" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag>‐treated cells. Error bars represent the mean ± s.e.m. (n = 3 experiments).<graphic href="https://api.sourcedata.io/file.php?panel_id=2267"/></sd-panel><sd-panel panel_id="2268">B <sd-tag id="sdTag287" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag288" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag289" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> cells were treated with <sd-tag id="sdTag290" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> (<sd-tag id="sdTag291" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag>), 25 μM <sd-tag id="sdTag292" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A), 25 μM <sd-tag id="sdTag293" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> and 10 μM <sd-tag id="sdTag298" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="oligomycin" ext_ids="CHEBI:25675" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="oligomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">oligomycin</sd-tag> (anti A + oligo), or 20 μM <sd-tag id="sdTag294" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag> (<sd-tag id="sdTag295" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag>) for two hours, then fixed and <sd-tag id="sdTag296" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> for <sd-tag id="sdTag297" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (red). Scale bars, 50 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2268"/></sd-panel><sd-panel panel_id="2269">C Quantification of<sd-tag id="sdTag460" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDHE2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDHE2/E3</sd-tag> bp−positive/<sd-tag id="sdTag470" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative structures from (B); both the total number (white bars) and the number colocalizing with <sd-tag id="sdTag471" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag472" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (gray bars) are indicated. Error bars represent the mean ± s.e.m. P−values are given first for <sd-tag id="sdTag473" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−/<sd-tag id="sdTag474" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag475" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag>−positive <sd-tag id="sdTag476" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicles" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicles</sd-tag>, then for total <sd-tag id="sdTag477" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vesicle" ext_ids="GO:0031982" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vesicle" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vesicle</sd-tag> number (n = 42-60 cells in three experiments); ns, not significant; ***P < 0.001).<graphic href="https://api.sourcedata.io/file.php?panel_id=2269"/></sd-panel><sd-panel panel_id="2270">D <sd-tag id="sdTag314" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag315" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag316" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> cells were treated with <sd-tag id="sdTag317" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag> (<sd-tag id="sdTag318" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag>), 25 μM <sd-tag id="sdTag319" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A), 25 μM <sd-tag id="sdTag320" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> and 10 μM <sd-tag id="sdTag321" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="oligomycin" ext_ids="CHEBI:25675" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="oligomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">oligomycin</sd-tag> (anti A + oligo), or 20 μM <sd-tag id="sdTag322" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag> (<sd-tag id="sdTag323" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag>) for two hours, then fixed and <sd-tag id="sdTag324" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> for <sd-tag id="sdTag461" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3</sd-tag> bp (red) and <sd-tag id="sdTag326" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (blue). A merged image of both <sd-tag id="sdTag437" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondrial" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondrial</sd-tag> markers (mito) is shown at the far right. Arrows indicate <sd-tag id="sdTag462" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E2/E3" ext_ids="P09622///P10515" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="DLD///DLAT" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">PDH E2/E3</sd-tag> bp−positive/<sd-tag id="sdTag328" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>−negative structures colocalizing with <sd-tag id="sdTag329" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag330" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> (green). Scale bars, 20 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2270"/></sd-panel></fig><fig id="1131"><title></title><label>Figure 7</label><graphic href="https://api.sourcedata.io/file.php?figure_id=1131"/><sd-panel panel_id="2271">A Representative <sd-tag id="sdTag331" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag> of whole‐cell lysates from <sd-tag id="sdTag332" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag333" source="sdapi" category="entity" entity_type="protein" role="component" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> and <sd-tag id="sdTag334" source="sdapi" category="entity" entity_type="protein" role="component" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag335" source="sdapi" category="entity" entity_type="gene" role="intervention" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> cells treated with <sd-tag id="sdTag336" source="sdapi" category="entity" entity_type="molecule" role="component" text="DMSO" ext_ids="CHEBI:28262" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="dimethyl sulfoxide" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DMSO</sd-tag>, 25 μM <sd-tag id="sdTag337" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> (anti A), 25 μM <sd-tag id="sdTag338" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="antimycin A" ext_ids="CHEBI:22584" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="antimycin A" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">antimycin A</sd-tag> with 10 μM <sd-tag id="sdTag339" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="oligomycin" ext_ids="CHEBI:25675" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="oligomycin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">oligomycin</sd-tag> (anti A + oligo), or 20 μM <sd-tag id="sdTag340" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="CCCP" ext_ids="CHEBI:3259" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="CCCP" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">CCCP</sd-tag> for the indicated time period.<graphic href="https://api.sourcedata.io/file.php?panel_id=2271"/></sd-panel><sd-panel panel_id="2272">B Quantification of <sd-tag id="sdTag463" source="sdapi" category="entity" entity_type="protein" role="assayed" text="PDH E1a" ext_ids="P08559" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PDHA1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">PDH E1a</sd-tag> signal intensity relative to that of <sd-tag id="sdTag411" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="actin" ext_ids="P60709" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ACTB" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">actin</sd-tag> in immunoblots from (A). Error bars represent the mean ± s.e.m. (n = 3 experiments).<graphic href="https://api.sourcedata.io/file.php?panel_id=2272"/></sd-panel><sd-panel panel_id="2273">C Quantification of <sd-tag id="sdTag452" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondrial" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondrial</sd-tag> clearance in <sd-tag id="sdTag363" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag364" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>−<sd-tag id="sdTag365" source="sdapi" category="entity" entity_type="gene" role="component" text="parkin" ext_ids="5071" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">parkin</sd-tag> cells treated as in (A), fixed and <sd-tag id="sdTag366" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> for <sd-tag id="sdTag367" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag>. Data are shown as percentage of cells containing <sd-tag id="sdTag368" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>, by <sd-tag id="sdTag369" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> staining, visualized by <sd-tag id="sdTag370" source="sdapi" category="assay" entity_type="" role="" text="fluorescence microscopy" ext_ids="BAO_0000450" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="fluorescence microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">fluorescence microscopy</sd-tag>. Error bars represent the mean ± s.e.m. (n = 3 experiments, with at least 85 cells quantified per condition, per experiment).<graphic href="https://api.sourcedata.io/file.php?panel_id=2273"/></sd-panel><sd-panel panel_id="2274">D <sd-tag id="sdTag380" source="sdapi" category="entity" entity_type="cell_line" role="component" text="U2OS" ext_ids="CVCL_0042" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="U2OS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">U2OS</sd-tag>:<sd-tag id="sdTag381" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>‐<sd-tag id="sdTag382" source="sdapi" category="entity" entity_type="protein" role="assayed" text="parkin" ext_ids="O60260" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="PARK2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">parkin</sd-tag> cells were treated as in (A) for the indicated time period, then fixed and <sd-tag id="sdTag383" source="sdapi" category="assay" entity_type="" role="" text="immunostained" ext_ids="BAO_0002426" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunofluorescent labeling" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunostained</sd-tag> for <sd-tag id="sdTag384" source="sdapi" category="entity" entity_type="protein" role="assayed" text="TOM20" ext_ids="Q15388" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TOMM20" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TOM20</sd-tag> (red). Cell boundaries are delineated in single‐channel images. Scale bars, 20 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=2274"/></sd-panel></fig></article>