Overexpression of cofilin rescues phenotypes of F-actin accumulations, shrinking protrusions, and impaired cell migration and invasion in WDR1 knockdown cells. A: shWDR1-treated A549 cells were transfected with pCDNA-Flag-cofilin plasmid, and transfection efficiency of Flag-cofilin was determined by western blotting. Enforced overexpression of cofilin declined the increased level of actin induced by WDR1 knockdown. Quantitation of the relative proteins to the corresponding control was also shown. B: Double immunostaining for F-actin and Flag-cofilin, using the antibody of Flag. Overexpression of Flag-cofilin in shWDR1 cells rescued phenotypes of F-actin accumulations and cell morphological disruptions. C-D: Matrigel invasion assay (C) and wound healing assay (D) in A549 and H1299 cells. Cell migratory rate and invasive ability were partly recovered after enforced overexpression of cofilin. Data are expressed as means ± SEM. **P < 0.01, and ***P < 0.001. Scale bar = 10μm.