Figure 4

PAK1 siRNA enhances the inhibitory effect of EOPK on the cell proliferation and migration. (A) HCT116 cells were transfected with PAK1-siRNA for 48 h and were incubated in the presence or absence of EOPK (100 μg/ml) for 24 h. Cell lysates were prepared and subjected to Western blotting to determine the expression of Cyclin D1, CDK4, CDK6 and β-actin. (B) HCT116 cells were transfected with PAK1-siRNA for 24 h and treated with EOPK (100 μg/ml) for 5 days. The cells were resolved in 70% ethanol after washing with distilled water, and crystal violet absorbance was read using a microplate reader. Data represent mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with control. ### p < 0.001 compared with control-siRNA and PAK1-siRNA. (C) Cell migration was assessed by a wound healing assay. The number of cells migrated into the scratched area was calculated. Data are shown as the mean ± SD of three independent experiments. *** p < 0.001 compared with untreated control. ### p < 0.001 compared with control-siRNA and PAK1-siRNA. (D) Fixed cells were stained with rhodamine phalloidin and DAPI, and imaged using a DV Elite™ Imaging System (Deltavision, Applied Precision, Inc., USA). Arrow indicates lamellipodia. Arrow indicates lamellipodia.