ADF and cofilin suppression affects MTLn3 cell polarization and pADF levels after EGF stimulation. A. MTLn3 cells were stimulated with 5 nM EGF for 0, 60 and 180 s and subdivided as polarized or non-polarized. B. MTLn3 cells were scored for percentage of polarized cells. ADF KD and cofilin KD cells are more polarized than control cells at the start of EGF treatment, but polarize less in response to EGF over the 180 s of EGF stimulation. C. Representative Western blot for pADF and pCofilin in control and EGF-treated MTLn3 cells after 180 s of EGF stimulation. Lane 1–2, control uninfected; lane 3–4 control infected; lane 5–6, ADF KD; Lane 7–8, cofilin KD. The experiment was repeated three times for 0, 60 and 180 s of EGF stimulation and the corresponding quantification is shown in (D). D. Relative levels of pCofilin and pADF after 60 and 180 s of EGF stimulation compared to their levels of the same treatment at 0 s obtained from densitometry of immunoblots. E. Relative levels of pCofilin in ADF KD cells and pADF in cofilin KD cells compared to controls (set at 1) obtained from densitometry of immunoblots. * p < 0.05, *** p < 0.001 versus control at the same time point. ∑ p < 0.05, ∑∑ p < 0.01, ∑∑∑ p < 0.001 versus same treatment at 0 s. Scale bar: 10 μm.