Figure 6.
YB-1 regulates filopodia formation through modulating Rho GTPases activity and reorganization of the actin cytoskeleton in HuH-7R cells. (A) YB-1 knockdown induced filopodia regression. F-actin was stained with phalloidin-conjugated Alexa Fluor 488 (green). DNA was counterstained with DAPI (blue). The number and length of filopodia per cell were calculated from 20 cells in each group (*** p < 0.001, Scale bars = 20 µm). (B) G-actin and F-actin were visualized via immunofluorescence staining with Alexa Fluor 488 phalloidin (F-actin, green) and DNase I-conjugated Alexa Fluor 594 (G-actin, red). The F-actin to G-actin ratio per cell was calculated from 50 cells in each group (*** p < 0.001, Scale bar = 50 μm). (C) Rho-family-GTPase activity was examined using a small GTPase family pull-down assay. Total cell lysates were immunoprecipitated and analyzed by immunoblotting with specific antibodies. Active cdc42 was quantified and normalized to total cdc42. Error bars indicate mean ± SEM, and p values were calculated with the t test (* p < 0.05).