Fig. 3

ELK3-CYFIP2 axis regulates metastatic nature of TNBCs by modulating flopodia protrusion. A Immunoblot analysis confrms the activity of siRNA targeting CYFIP2 (siCYFIP2) in ELK3KD MDA-MB-231 and Hs578T cells. ELK3KD TNBC cells transfected with a non-specifc siRNA (siNS) or siCYFIP2. Relative band intensity of ELK3 and CYFIP2. Data are presented as the mean ± SD. B Filopodia formation was observed after staining with DAPI and phalloidin. Actin accumulation of flopodia formation was visualized using fuorescence microscopy; representative protrusions are indicated by red arrows. Scale bar, 20 μm. C The number of flopodia per cell were quantifed, and is presented as individual dots. (MDA-MB-231 cells, n = 30, 30, and 26, respectively, Hs578T cells, n = 14, 30, and 30, respectively.) D The length of flopodia are presented in a graph. (MDA-MB-231 cells, n = 30, 30, and 26, respectively, Hs578T cells, n = 16, 30, and 30, respectively.) Data are presented as the SEM. E–F Representative images showing migration and adhesion of the indicated cells. Scale bar, 200 μm. All data were derived from at least three independent biological experiments. Data are presented as the mean ± SD. Control (Cont) = sh control of MDA-MB-231 or Hs578T cells; ELK3KD = ELK3KD of MDA-MB-231 or Hs578T cells. NS indicates no statistical signifcance. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001