FIGURE 1:

PDGF-induced CDR formation is affected by RhoG silencing. (A) A7r5 cells were transfected with siRNA against RhoG (RhoG KD) or with a nontargeting siRNA (CTRL). After 72 h, cells were serum starved for 2 h and stimulated with PDGF-BB (20 ng/ml) for 1, 2.5, and 5 min. Cells were fixed and processed for immunofluorescence using cortactin (red) as marker of dorsal ruffles, Alexa 488–phalloidin (green) to stain F-actin, and Hoechst (blue) to visualize nuclei. Representative images of the 2.5-min time point. Scale bar, 20 μm. (B) KD efficiency was tested for each experiment by immunoblot in cell lysates 72 h after transfection. (C) Number of cells with CDRs expressed as percentage of cells that stained positive for at least one CDR. (D) Average number of CDRs per cell (in cells positive for CDRs). (E) Average CDR area. Black bars, CTRL cells; gray bars, RhoG KD. Results are shown as mean ± SEM from three independent experiments (≥100 cells per condition/experiment).