Abstract
Western blot analysis for Tubulin (cytoplasmic marker), Lamin A/C (nuclear marker), and Calnexin (ER marker) showing the purity of each fraction prepared from siCtrl-, siAGO1-or siAGO2-treated MDA-MB-453 cells. siRNAs specific for AGO1 or AGO2 were transfected into cells at 25 nM using Lipofectamine RNAiMAX. Cells were harvested 3 days after transfection followed by subcellular fractionation. Equal amount of proteins (25 μg) from each fraction was analyzed by SDS-PAGE (4-20% TGX gels).

Western blot data for AGO1 and AGO2 in the whole cell and the cytoplasmic and the nuclear fractions after siAGO1 or siAGO2 treatment (upper) and the quantitation of AGO1 or AGO2 protein levels (lower, n=3). siCtrl, siAGO1, and siAGO2 were transfected into MDA-MB-453 cells at 25 nM. Equal amount of proteins (25 μg) from each fraction were analyzed by SDS-PAGE. Error bars are SD. *p<0.05, **p<0.01, and ***p<0.001 relative to siCtrl treatment (paired t-test). ) and a pool (-2/-3) (25 nM) for AGO1 knockdown. Two days after transfection, cells were harvested using TRIzol and total RNAs were isolated. RT reaction was performed using High Capacity cDNA Reverse Transcription Kit. qPCR was performed using iTaq supermix or iTaq SYBR supermix (Biorad) and primers specific for AGO1 mRNA (Hs00201864_m1) or 18S RNA (Forward: 5'-GACCAGAGCGAAAGCATTTG-3', Reverse: 5'-TCGGAACTACGACGGTATCT-3'). n=2. (B) Western blot analysis for Tubulin (cytoplasmic marker), Lamin A/C (nuclear marker), and Calnexin (ER marker) showing the purity of each fraction prepared from siCtrl-or siAGO1-b-treated T47D cells. (C) Western blot data showing levels of Dicer, TNRC6A, AGO1, AGO2, and TRBP proteins in the whole cell and the cytoplasmic and the nuclear fractions after siCtrl or siAGO1-b treatment. siCtrl and siAGO1-b were transfected into cells at 25 nM using Lipofectamine RNAiMAX. Cells were harvested 3 days after transfection followed by subcellular fractionation. Equal amount of proteins (25 μg) from each fraction was analyzed by SDS-PAGE (4-20% TGX gels). Data were representative from two independent experiments. 

{'paper_id': '11303449', 'title': 'Reduced Expression of Argonaute 1, Argonaute 2, and TRBP Changes Levels and Intracellular Distribution of RNAi Factors', 's2_url': 'https://api.semanticscholar.org/CorpusID:11303449', 'source': 'S2ORC'}