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<article doi="10.1002/eji.200323730" abstract="Biochemical and functional studies have demonstrated major histocompatibility complex (MHC) class II-restricted presentation of peptides derived from cytosolic proteins, but the underlying processing and presentation pathways have remained elusive. Here we show that endogenous presentation of an epitope derived from the cytosolic protein neomycin phosphotransferase II (NeoR) on MHC class II is mediated by autophagy. This presentation pathway involves the sequestration of NeoR into autophagosomes, and subsequent delivery into the lytic compartment. These results identify endosomes/lysosomes as the processing compartment for cytosolic antigens and furthermore link endogenous antigen presentation on MHC class II with the process of cellular protein turnover by autophagy."><fig id="710"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=710"/><sd-panel panel_id="980"><p><strong>(A)</strong> The <sd-tag id="sdTag1" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cell" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cell</sd-tag> clone <sd-tag id="sdTag110" source="sdapi" category="entity" entity_type="cell_type" role="component" text="20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">20-4/A4</sd-tag> recognizes an epitope derived from the <sd-tag id="sdTag5" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein in the context of <sd-tag id="sdTag8" source="sdapi" category="entity" entity_type="subcellular" role="component" text="HLA‐DP3" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP3</sd-tag>. <sd-tag id="sdTag13" source="sdapi" category="entity" entity_type="subcellular" role="component" text="MHC class II" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MHC class II</sd-tag> expression on <sd-tag id="sdTag10" source="sdapi" category="entity" entity_type="cell" role="experiment" text="RCC1.24" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RCC1.24</sd-tag>‐<sd-tag id="sdTag270" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> cells following <sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="protein" role="intervention" text="IFN‐γ" ext_ids="P01579" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="IFNG" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">IFN‐γ</sd-tag> treatment confers recognition by <sd-tag id="sdTag283" source="sdapi" category="entity" entity_type="cell_type" role="component" text="20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">20-4/A4</sd-tag>. This recognition is almost completely blocked by the <sd-tag id="sdTag272" source="sdapi" category="entity" entity_type="subcellular" role="component" text="HLA‐DP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP</sd-tag>‐specific antibody <sd-tag id="sdTag550">B7/21</sd-tag>. <sd-tag id="sdTag50">EBV‐B</sd-tag> cells transfected with <sd-tag id="sdTag51">pINCO</sd-tag>‐<sd-tag id="sdTag7" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> are able to present endogenous <sd-tag id="sdTag273" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> on <sd-tag id="sdTag14" source="sdapi" category="entity" entity_type="subcellular" role="component" text="MHC class II" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MHC class II</sd-tag>. As exemplified by the cell lines <sd-tag id="sdTag52">EBV‐B1.11</sd-tag> (<sd-tag id="sdTag16" source="sdapi" category="entity" entity_type="subcellular" role="component" text="HLA‐DP3" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP3</sd-tag>) and <sd-tag id="sdTag53">EBV‐B1.25</sd-tag> (HLA‐DP4), recognition by <sd-tag id="sdTag284" source="sdapi" category="entity" entity_type="cell_type" role="component" text="20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">20-4/A4</sd-tag> is dependent on <sd-tag id="sdTag276" source="sdapi" category="entity" entity_type="subcellular" role="component" text="HLA‐DP3" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP3</sd-tag> expression. Bacterially expressed <sd-tag id="sdTag274" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein and a synthetic peptide encompassing amino acids <sd-tag id="sdTag275" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag><sub>216-229</sub> served as controls.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=980"/></sd-panel><sd-panel panel_id="981"><b>(B)</b> <sd-tag id="sdTag55" source="sdapi" category="assay" entity_type="" role="" text="Cross-presentation experiments" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Cross-presentation experiments</sd-tag> to exclude release of <sd-tag id="sdTag38" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> and re‐uptake as exogenous protein. <sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.25" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.25</sd-tag> cells transfected with the <sd-tag id="sdTag56" source="sdapi" category="entity" entity_type="gene" role="intervention" text="neoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls=""><i>neoR</i></sd-tag> expression plasmid <sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag>, were co‐cultured for 24 h with the same number of untransfected <sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells, or vice versa. Then, <sd-tag id="sdTag57" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cells" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cells</sd-tag> were added and co‐cultured with the cells for additional 24 h. To test for <sd-tag id="sdTag277" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> release by <i><sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="gene" role="intervention" text="neoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neoR</sd-tag></i>‐transfected <sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells, total <sd-pretag id="sdPretag14253982055630" parent-tag-id="147" class="component subcellular">supernatant</sd-pretag> (200 μl) of 1×10<sup>5</sup> <sd-tag id="sdTag45" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells was harvested 48 h post transfection by centrifugation, and was used as culture media for 1×10<sup>5</sup> untransfected <sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells. After 48 h, 1×10<sup>5</sup> <sd-tag id="sdTag111" source="sdapi" category="entity" entity_type="cell_type" role="component" text="20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">20-4/A4</sd-tag> <sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cells" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cells</sd-tag> were added and <sd-tag id="sdTag54" source="sdapi" category="entity" entity_type="protein" role="assayed" text="GM‐CSF" ext_ids="P04141" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CSF2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">GM‐CSF</sd-tag> concentration determined after additional 24 h.<graphic href="https://api.sourcedata.io/file.php?panel_id=981"/></sd-panel></fig><fig id="711"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=711"/><sd-panel panel_id="982"><p>(A), <sd-tag id="sdTag60" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag61" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> transfected <sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="cell_type" role="experiment" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells (B), and as control <sd-tag id="sdTag63" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="gene" role="component" text="Tyrosinase" ext_ids="7299" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TYR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Tyrosinase</sd-tag>‐transfected <sd-tag id="sdTag278" source="sdapi" category="entity" entity_type="cell_type" role="experiment" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells (C), were either left untreated or treated with 5 μM <sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="lactacystin" ext_ids="CHEBI:52722" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="lactacystin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">lactacystin</sd-tag> or 50 μM <sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="AAF‐CMK" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">AAF‐CMK</sd-tag> for 20 h. Following fixation with 0.5% paraformaldehyde, cells were incubated with <sd-tag id="sdTag65" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cells" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cells</sd-tag>, and 24 h later <sd-tag id="sdTag67" source="sdapi" category="entity" entity_type="protein" role="assayed" text="GM‐CSF" ext_ids="P04141" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CSF2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">GM‐CSF</sd-tag> release was determined by <sd-tag id="sdTag72" source="sdapi" category="assay" entity_type="" role="" text="ELISA" ext_ids="BAO_0000134" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="ELISA" 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">ELISA</sd-tag>.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=982"/></sd-panel><sd-panel panel_id="983"><p>(D) <sd-tag id="sdTag84" source="sdapi" category="assay" entity_type="" role="" text="FACS analysis" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">FACS analysis</sd-tag> of <sd-tag id="sdTag77" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells (open histograms) and <sd-tag id="sdTag279" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells stably transfected with the <sd-tag id="sdTag85" source="sdapi" category="entity" entity_type="gene" role="intervention" text="ICP47" ext_ids="1487353" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="US12" ext_tax_ids="10310" ext_tax_names="Human alphaherpesvirus 2" ext_urls="http://www.ncbi.nlm.nih.gov/gene/"><i>ICP47</i></sd-tag> gene (shaded histograms). Inhibition of <sd-tag id="sdTag86" source="sdapi" category="entity" entity_type="protein" role="component" text="TAP" ext_ids="Q03518" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TAP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">TAP</sd-tag> by <sd-tag id="sdTag88" source="sdapi" category="entity" entity_type="gene" role="intervention" text="ICP47" ext_ids="1487353" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="US12" ext_tax_ids="10310" ext_tax_names="Human alphaherpesvirus 2" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">ICP47</sd-tag> down‐regulates <sd-tag id="sdTag89" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA class I" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA class I</sd-tag> but not <sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA‐DP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP</sd-tag> expression. An isotype‐matched antibody was used as control (thin‐lined histogram). <sd-tag id="sdTag280" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag>‐<sd-tag id="sdTag282" source="sdapi" category="entity" entity_type="gene" role="intervention" text="ICP47" ext_ids="1487353" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="US12" ext_tax_ids="10310" ext_tax_names="Human alphaherpesvirus 2" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">ICP47</sd-tag> cells were transfected with <sd-tag id="sdTag80" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag79" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> (E), <sd-tag id="sdTag82" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag81" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Tyrosinase" ext_ids="7299" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TYR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Tyrosinase</sd-tag> (F), or mock transfected (<sd-tag id="sdTag281" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>) and <sd-tag id="sdTag90" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="MHC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MHC</sd-tag>‐restricted presentation monitored with <sd-tag id="sdTag83" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cells" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cells</sd-tag>.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=983"/></sd-panel></fig><fig id="712"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=712"/><sd-panel panel_id="984"><p>(A) <sd-tag id="sdTag92" source="sdapi" category="entity" entity_type="cell" role="experiment" text="RCC1.24" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RCC1.24</sd-tag>‐<sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> incubated with <sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="protein" role="component" text="IFN‐γ" ext_ids="P01579" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="IFNG" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">IFN‐γ</sd-tag> were either left untreated (w/o) or treated with <sd-tag id="sdTag103" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="chloroquine" ext_ids="CHEBI:3638" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="chloroquine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">chloroquine</sd-tag> or <sd-tag id="sdTag104" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for 24 h to block lysosomal processing. Subsequently cells were fixed with 0.5% paraformaldehyde and tested with <sd-tag id="sdTag105" source="sdapi" category="entity" entity_type="cell_type" role="component" text="20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">20-4/A4</sd-tag> in a <sd-tag id="sdTag96" source="sdapi" category="entity" entity_type="protein" role="assayed" text="GM‐CSF" ext_ids="P04141" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CSF2" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">GM‐CSF</sd-tag> release assay. <sd-tag id="sdTag97" source="sdapi" category="entity" entity_type="cell_type" role="experiment" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells transfected with <sd-tag id="sdTag98" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag99" source="sdapi" category="entity" entity_type="gene" role="intervention" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> (B) or <sd-tag id="sdTag100" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag101" source="sdapi" category="entity" entity_type="gene" role="component" text="Tyrosinase" ext_ids="7299" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TYR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Tyrosinase</sd-tag> (C) were tested likewise. Both substances abrogated recognition of <sd-tag id="sdTag102" source="sdapi" category="entity" entity_type="gene" role="intervention" text="neoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neoR</sd-tag>‐transfected cells, while <sd-tag id="sdTag106" source="sdapi" category="entity" entity_type="subcellular" role="component" text="HLA‐A2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐A2</sd-tag>‐restricted presentation of the <sd-tag id="sdTag107" source="sdapi" category="entity" entity_type="protein" role="component" text="tyrosinase" ext_ids="P14679" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TYR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">tyrosinase</sd-tag> peptide was not affected</p><graphic href="https://api.sourcedata.io/file.php?panel_id=984"/></sd-panel><sd-panel panel_id="985"><p><strong>(D)</strong> ) <sd-tag id="sdTag112" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells were incubated with 200 ng/ml of recombinant <sd-tag id="sdTag113" source="sdapi" category="entity" entity_type="protein" role="component" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein and treated with <sd-tag id="sdTag114" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> or <sd-tag id="sdTag115" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="chloroquine" ext_ids="CHEBI:3638" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="chloroquine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">chloroquine</sd-tag> as described in (A). Presentation of exogenous <sd-tag id="sdTag118" source="sdapi" category="entity" entity_type="protein" role="component" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> on <sd-tag id="sdTag117" source="sdapi" category="entity" entity_type="subcellular" role="component" text="HLA‐DP3" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP3</sd-tag> is blocked by both substances as well, indicating that <sd-tag id="sdTag285" source="sdapi" category="entity" entity_type="protein" role="component" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> presentation is dependent on lysosomal processing.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=985"/></sd-panel></fig><fig id="713"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=713"/><sd-panel panel_id="986"><p><strong>(A)</strong> <sd-tag id="sdTag130" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293T</sd-tag> cells transfected with <sd-tag id="sdTag122" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> were harvested 6 and 96 h post transfection and <sd-tag id="sdTag286" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> localization within various cell fractions determined by <sd-tag id="sdTag137" source="sdapi" category="assay" entity_type="" role="" text="Western blot" 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">Western blot</sd-tag>. In addition, <sd-tag id="sdTag148" source="sdapi" category="entity" entity_type="undefined" role="experiment" text="supernatant" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">supernatant</sd-tag> of <sd-tag id="sdTag129" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>‐<sd-tag id="sdTag125" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag> transfected <sd-tag id="sdTag134" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293T</sd-tag> cells (96 h post transfection) was included to test for release of <sd-tag id="sdTag128" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> into the culture <sd-tag id="sdTag289" source="sdapi" category="entity" entity_type="undefined" role="experiment" text="supernatant" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">supernatant</sd-tag>. At 6 h post transfection, <sd-tag id="sdTag287" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein becomes detectable in the <sd-tag id="sdTag138" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="cytoplasm" ext_ids="GO:0005737" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="cytoplasm" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">cytoplasm</sd-tag>, while at 96 h post transfection a substantial amount of <sd-tag id="sdTag288" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein is also localized in the <sd-tag id="sdTag139" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="endosomal/lysosomal" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">endosomal/lysosomal</sd-tag> fraction. For further characterization of the various fractions, blots were stripped and re‐probed with antibodies specific for the <sd-pretag id="sdPretag14253998394220" parent-tag-id="206" class="component subcellular">cytoplasmic</sd-pretag> protein <sd-tag id="sdTag140" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LDH" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">LDH</sd-tag>, the nuclear protein <sd-tag id="sdTag141" source="sdapi" category="entity" entity_type="protein" role="assayed" text="c‐myc" ext_ids="P01106" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MYC" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">c‐myc</sd-tag>, early endosomes (<sd-tag id="sdTag142" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Rab‐5" ext_ids="P61020///P20339" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="RAB5B///RAB5A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Rab‐5</sd-tag>), late endosomes and lysosomes (<sd-tag id="sdTag145" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Lamp‐1" ext_ids="P11279" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="LAMP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Lamp‐1</sd-tag>) and <sd-tag id="sdTag149" 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> (<sd-tag id="sdTag144" source="sdapi" category="entity" entity_type="protein" role="assayed" text="cytochrome c" ext_ids="P99999" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CYCS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">cytochrome c</sd-tag>), which co‐purify with <sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="endosomes/lysosomes" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">endosomes/lysosomes</sd-tag>.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=986"/></sd-panel><sd-panel panel_id="987"><b>(B)</b> <sd-tag id="sdTag166" source="sdapi" category="entity" entity_type="subcellular" role="component" text="Endosomal/lysosomal" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">Endosomal/lysosomal</sd-tag> fractions were prepared from <sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag155" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>‐transfected cells 96 h post transfection as in (A). To prevent lysosomal degradation of <sd-tag id="sdTag290" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>, half of the cells were treated with <sd-tag id="sdTag156" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> for the last 48 h. These fractions were separated on a 27% Percoll gradient and eight different fractions collected. Successful separation into fractions with increasing density was verified by <sd-tag id="sdTag158" source="sdapi" category="assay" entity_type="" role="" text="Western blot" 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">Western blot</sd-tag> analysis with antibodies directed against <sd-tag id="sdTag159" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Lamp‐1" ext_ids="P11279" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="LAMP1" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">Lamp‐1</sd-tag>, <sd-tag id="sdTag160" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Rab‐5" ext_ids="P61020///P20339" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="RAB5B///RAB5A" ext_tax_ids="9606///9606" ext_tax_names="Homo sapiens///Homo sapiens" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Rab‐5</sd-tag> and <sd-tag id="sdTag161" source="sdapi" category="entity" entity_type="protein" role="assayed" text="cytochrome c" ext_ids="P99999" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="CYCS" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">cytochrome c</sd-tag>. Inhibition of <sd-tag id="sdTag164" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> degradation by <sd-tag id="sdTag163" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> causes accumulation of <sd-tag id="sdTag291" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein in the late <sd-tag id="sdTag165" source="sdapi" category="entity" entity_type="subcellular" role="component" text="endosomal/lysosomal" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">endosomal/lysosomal</sd-tag> fractions 7 and 8.<graphic href="https://api.sourcedata.io/file.php?panel_id=987"/></sd-panel><sd-panel panel_id="988"><b>(C)</b> To determine where <sd-tag id="sdTag167" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> gains access to the vacuolar system, <sd-tag id="sdTag168" source="sdapi" category="entity" entity_type="cell_line" role="component" text="293T" ext_ids="CVCL_0063" norm_text="" ext_dbs="CVCL" in_caption="True" ext_names="HEK293T" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://identifiers.org/cellosaurus:">293T</sd-tag> cells were transfected with <sd-tag id="sdTag169" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag172" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>, treated with <sd-tag id="sdTag171" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> to prevent <sd-tag id="sdTag174" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> degradation, and the <sd-tag id="sdTag173" source="sdapi" category="entity" entity_type="subcellular" role="component" text="endosomal/lysosomal" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">endosomal/lysosomal</sd-tag> fractions prepared at various time points post transfection and separated by <sd-tag id="sdTag179" source="sdapi" category="assay" entity_type="" role="" text="Percoll gradient centrifugation" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Percoll gradient centrifugation</sd-tag>. Starting at 16 h post transfection, <sd-tag id="sdTag292" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> becomes detectable in the Rab‐5‐positive fraction 5, and then spreads into late <sd-tag id="sdTag176" source="sdapi" category="entity" entity_type="subcellular" role="component" text="endosomal/lysosomal" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">endosomal/lysosomal</sd-tag> fractions. At 26 h post transfection an almost identical <sd-tag id="sdTag177" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> distribution pattern is observed in cells treated with or without BfA.<graphic href="https://api.sourcedata.io/file.php?panel_id=988"/></sd-panel></fig><fig id="714"><title></title><label>Figure 5</label><graphic href="https://api.sourcedata.io/file.php?figure_id=714"/><sd-panel panel_id="989"><b>(A)</b> <sd-tag id="sdTag183" source="sdapi" category="entity" entity_type="cell" role="component" text="RCC" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RCC</sd-tag> were transfected with the <sd-tag id="sdTag182" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>‐<sd-tag id="sdTag184" 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> expression construct and subcellular localization of the fusion protein monitored by <sd-tag id="sdTag187" source="sdapi" category="assay" entity_type="" role="" text="UV fluorescence" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">UV fluorescence</sd-tag>. As compared to untreated cells, cells treated with <sd-pretag id="sdPretag14253992910590" parent-tag-id="191" class="component molecule">3‐MA</sd-pretag> show a much brighter <sd-tag id="sdTag293" source="sdapi" category="assay" entity_type="" role="" text="UV fluorescence" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">UV fluorescence</sd-tag>.<graphic href="https://api.sourcedata.io/file.php?panel_id=989"/></sd-panel><sd-panel panel_id="990"><b>(B)</b> Semiquantitative analysis of fluorescence intensity of cells transfected with the <sd-tag id="sdTag188" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>‐<sd-tag id="sdTag189" 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> fusion construct. Compared to untreated cells, cells treated for 24 h with 7.5 mM <sd-tag id="sdTag191" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag> show an about threefold increase in mean fluorescence intensity.<graphic href="https://api.sourcedata.io/file.php?panel_id=990"/></sd-panel><sd-panel panel_id="991"><b>(C)</b> <sd-tag id="sdTag195" source="sdapi" category="assay" entity_type="" role="" text="Western blot" 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">Western blot</sd-tag> analysis of <i><sd-tag id="sdTag196" source="sdapi" category="entity" entity_type="gene" role="component" text="neoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">neoR</sd-tag></i>‐transfected cells. Treatment of cells with 7.5 mM <sd-tag id="sdTag197" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag> causes a dramatic increase in <sd-tag id="sdTag198" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein levels in only 12 h. The ER‐resident protein <sd-tag id="sdTag200" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="BiP" ext_ids="P11021" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HSPA5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BiP</sd-tag> was used as gel loading control.<graphic href="https://api.sourcedata.io/file.php?panel_id=991"/></sd-panel><sd-panel panel_id="992"><b>(D)</b> This increase in total <sd-tag id="sdTag202" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein in <sd-tag id="sdTag203" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag>‐treated cells is associated with an increase in <sd-tag id="sdTag206" source="sdapi" category="entity" entity_type="subcellular" role="component" text="cytoplasmic" ext_ids="GO:0005737" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="cytoplasm" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">cytoplasmic</sd-tag>, and a decrease in <sd-tag id="sdTag204" source="sdapi" category="entity" entity_type="subcellular" role="component" text="endosomal/lysosomal" ext_ids="GO:0005764///GO:0005768" norm_text="" ext_dbs="Gene Ontology///Gene Ontology" in_caption="True" ext_names="lysosome///endosome" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term////http://amigo.geneontology.org/amigo/term/">endosomal/lysosomal</sd-tag> <sd-tag id="sdTag205" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein levels.<graphic href="https://api.sourcedata.io/file.php?panel_id=992"/></sd-panel><sd-panel panel_id="993"><b>(E)</b> <sd-tag id="sdTag212" source="sdapi" category="assay" entity_type="" role="" text="Western blot" 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">Western blot</sd-tag> analysis of whole cell lysates prepared from <i><sd-tag id="sdTag213" source="sdapi" category="entity" entity_type="gene" role="component" text="neoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">neoR</sd-tag></i>‐transfected cells. Inhibition of protein synthesis by cycloheximide treatment leads to a rapid decrease in <sd-tag id="sdTag216" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein levels. Simultaneous treatment of cells with <sd-tag id="sdTag215" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag> prevents <sd-tag id="sdTag294" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> degradation. <sd-tag id="sdTag217" source="sdapi" category="entity" entity_type="protein" role="normalizing" text="BiP" ext_ids="P11021" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="HSPA5" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">BiP</sd-tag> was again used as a gel loading control.<graphic href="https://api.sourcedata.io/file.php?panel_id=993"/></sd-panel></fig><fig id="715"><title></title><label>Figure 6</label><graphic href="https://api.sourcedata.io/file.php?figure_id=715"/><sd-panel panel_id="994"><b>(A)</b> <sd-tag id="sdTag222" source="sdapi" category="entity" entity_type="cell" role="component" text="RCC1.24" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RCC1.24</sd-tag>‐<sd-tag id="sdTag223" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> (endogenous), and <sd-tag id="sdTag295" source="sdapi" category="entity" entity_type="cell" role="component" text="RCC1.24" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">RCC1.24</sd-tag> cells to which <sd-tag id="sdTag225" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag> protein had been added (exogenous), were treated with different concentrations of <sd-tag id="sdTag224" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag> and cell surface presentation of the <sd-tag id="sdTag296" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">NeoR</sd-tag>‐derived peptide on <sd-tag id="sdTag226" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA‐DP3" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP3</sd-tag> monitored with the <sd-tag id="sdTag228" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cell clone 20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cell clone 20-4/A4</sd-tag>‐specific <sd-tag id="sdTag297" source="sdapi" category="entity" entity_type="cell_type" role="component" text="T cell clone 20–4/A4" ext_ids="CL:0000084" norm_text="" ext_dbs="CL" in_caption="True" ext_names="T cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">T cell clone 20-4/A4</sd-tag>. Inhibition of autophagy by <sd-tag id="sdTag231" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag> causes an almost complete inhibition of endogenous antigen presentation, while presentation of exogenous <sd-tag id="sdTag232" source="sdapi" category="entity" entity_type="protein" role="intervention" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> is not affected.<graphic href="https://api.sourcedata.io/file.php?panel_id=994"/></sd-panel><sd-panel panel_id="995"><b>(B)</b> <sd-tag id="sdTag233" source="sdapi" category="entity" entity_type="cell_type" role="component" text="EBV‐B1.11" ext_ids="CL:0000236" norm_text="" ext_dbs="CL" in_caption="True" ext_names="B cell" ext_tax_ids="" ext_tax_names="" ext_urls=" https://identifiers.org/">EBV‐B1.11</sd-tag> cells transfected with <sd-tag id="sdTag234" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag236" source="sdapi" category="entity" entity_type="protein" role="component" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> (endogenous), or incubated with recombinant <sd-tag id="sdTag298" source="sdapi" category="entity" entity_type="protein" role="component" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag> protein (exogenous), were treated with the inhibitors of autophagy, <sd-tag id="sdTag241" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="wortmannin" ext_ids="CHEBI:52289" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="wortmannin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">wortmannin</sd-tag> and <sd-tag id="sdTag237" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="3‐MA" ext_ids="CHEBI:38635///CHEBI:25272" norm_text="" ext_dbs="ChEBI///ChEBI" in_caption="True" ext_names="3-methyladenine///methyladenine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=///http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">3‐MA</sd-tag>, and <sd-tag id="sdTag238" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA‐DP3" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP3</sd-tag>‐restricted presentation of the <sd-tag id="sdTag239" source="sdapi" category="entity" entity_type="protein" role="assayed" text="NeoR" ext_ids="Q76MW8" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="neoR" ext_tax_ids="474004" ext_tax_names="Dual-tagging gene trap vector pGT1" ext_urls="https://www.uniprot.org/uniprot/">NeoR</sd-tag>‐derived peptide determined. Inhibition of antigen presentation by <sd-tag id="sdTag240" source="sdapi" category="entity" entity_type="molecule" role="component" text="leupeptin" ext_ids="CHEBI:6426" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="Leupeptin" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">leupeptin</sd-tag> served as control.<graphic href="https://api.sourcedata.io/file.php?panel_id=995"/></sd-panel><sd-panel panel_id="996"><b>(C)</b> Presentation of the <sd-tag id="sdTag258" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA‐A2" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐A2</sd-tag>‐restricted <sd-tag id="sdTag257" source="sdapi" category="entity" entity_type="protein" role="assayed" text="tyrosinase" ext_ids="P14679" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="TYR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="https://www.uniprot.org/uniprot/">tyrosinase</sd-tag> peptide in <sd-pretag id="sdPretag14253685948180" parent-tag-id="22" class="component cell">EBV‐B1.11</sd-pretag> cells transfected with <sd-tag id="sdTag260" source="sdapi" category="entity" entity_type="gene" role="component" text="pINCO" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">pINCO</sd-tag>‐<sd-tag id="sdTag259" source="sdapi" category="entity" entity_type="gene" role="component" text="Tyrosinase" ext_ids="7299" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="TYR" ext_tax_ids="9606" ext_tax_names="Homo sapiens" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Tyrosinase</sd-tag> is not affected by these inhibitors of autophagy.<graphic href="https://api.sourcedata.io/file.php?panel_id=996"/></sd-panel><sd-panel panel_id="997"><p><sd-pretag id="sdPretag14253716979410" parent-tag-id="84" class="assay">FACS analysis</sd-pretag> of <sd-pretag id="sdPretag14255455279520" parent-tag-id="266">HLA‐class I</sd-pretag> and <sd-tag id="sdTag265" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA‐DP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐DP</sd-tag> expression on <sd-pretag id="sdPretag14253684415850" parent-tag-id="10" class="component cell">RCC1.24</sd-pretag> (D) and <sd-pretag id="sdPretag14253685958140" parent-tag-id="22" class="component cell">EBV‐B1.11</sd-pretag> cells (E) As compared to untreated cells (thick‐lined histogram), treatment of cells with 7.5 mM <sd-pretag id="sdPretag14253993005410" parent-tag-id="191" class="component molecule">3‐MA</sd-pretag> (shaded histogram) does not cause down‐regulation of <sd-tag id="sdTag266" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="HLA‐class I" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HLA‐class I</sd-tag> or <sd-pretag id="sdPretag14255456018070" parent-tag-id="265" class="assayed subcellular">HLA‐DP</sd-pretag>. Isotype control is shown as thin‐lined histogram.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=997"/></sd-panel></fig></article> |
