Figure 1

Subcellular fractionation of T47D cells to isolate whole cells, cytoplasmic fractions and nuclear fractions.
(A) Scheme of dsRNA transfection and subsequent subcellular fractionation. siRNAs specific for AGO1, AGO2, or TRBP and control dsRNA (siCtrl) were transfected into cells at 25 nM using Lipofectamine RNAiMAX. (B–D) 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-, siAGO2, or siTRBP-treated T47D cells. Equal amount of proteins (25 μg) from each fraction were analyzed by SDS-PAGE. Similar western blot data for MDA-MB-453 cells are presented in Figure S1.