Fig. 5

LIMK1 was required for PAK4-mediated NSCLC cell migration and invasion. a: The protein expression of LIMK1, p-LIMK1, cofilin, and p-cofilin in A549 and NCI-H520 cells transfected with si-PAK4 or control by western blotting. b: Immunofluorescent staining for PAK4 (green) and LIMK1 (red) in A549 cells (upper panels) or in NCI-H520 cells (lower panels). Nuclei were stained with Hoechst33258 (blue). c: A549 and NCI-H520 cell lysates were immunoprecipitated with PAK4 antibody (top panels) or LIMK1 antibody (bottom panels) and subjected to western blotting to ascertain LIMK1 and PAK4 interaction. d: In vitro kinase assay using purified activated PAK4 (S445N), kinase-defective PAK4 (K350M), PAK4 (WT), and LIMK1 protein. The amount of p-LIMK1, PAK4, and LIMK1 were measured using western blotting. e: Correlations between the protein level of PAK4 and the p-LIMK1 in human NSCLC tissues (n = 10). NC, negative control; IgG, immunoglobulin G