Figure 6

Rab5a regulates the activity of Rho GTPases and reorganization of actin cytoskeleton in HeLa cells. (A) The Control and Rab5a-siRNAs cells were fixed and observed by a scanning electron microscopy (SEM). ×5 000. Knockdown of Rab5a expression with both siRNAs (b, c) caused a significant reduction in the number and size of filopodia and lamellipodia formation compared with Control (a). (B) The Control and Rab5a-siRNAs cells were double labeled with DAPI and FITC-phalloidine. Scale bars indicate 20 μm. (C) Levels of active GTP-bound Rac1, Cdc42 and GFP-RhoA in Control and Rab5a-siRNAs cells were detected by GST-pull down assays, as described in the materials and methods. Total Rac1, Cdc42 and GFP-RhoA present in the lysates were analysed by immunoblotting with their specific antibody respectively.