EGF activates LIMK2 and Cofilin expression in MG63 cells through ROCK. (A) Phosphorylation of LIMK2 and Cofilin in MG63 cells treated with 10 ng/mL EGF for 2, 6 and 12 h, as detected by western blotting. Quantification of LIMK2 (B) and Cofilin (C) phosphorylation in MG63 cells treated with 10 ng/mL EGF for 2, 6 and 12 h; (D) Phosphorylation of LIMK2 and Cofilin pretreated with Y27632 for 2 h followed by the addition of EGF, as detected by western blotting. Quantification of Cofilin (E) and LIMK2 (F) phosphorylation in MG63 cells pretreated with Y27632 for 2 h followed by the addition of EGF; (G) Expression of LIMK2 and Cofilin in MG63 cells treated with LIMK2 siRNA and Cofilin1 siRNA, as detected by western blotting. siR-Ribo™ served as the negative control (NC); (H) Phalloidin staining of stress fibers and DAPI staining of MG63 cells treated with LIMK2 and Cofilin1 siRNAs. siR-Ribo™ served as the negative control, scale bar = 50 μm. * p < 0.05, ** p < 0.01.