VEGF-C led to activation of the actin-regulatory protein, moesin. (A-B) SiHa cells were treated with VEGF-C (100 ng/mL) for 24 or 48 h alone, or treated with VEGF-C for 48 h in the presence of Flt-4/IgG (100 ng/mL). Total cell amount of wild-type (Moesin) or Thr558-phosphorylated moesin (P-Moesin) or β-actin (Actin) are shown with western blot analysis. Densitometry values were adjusted to β-actin intensity and then normalized to expression from the control sample. ** = P < 0.01 vs. corresponding control. ## = P < 0.01 vs VEGF-C 48 h. (C) SiHa cells were transfected with scrambled siRNA (-S) or moesin targeted siRNA (-M) for 48 h. After that the level of moesin expression was detected by western blot as indicated. β-actin was used as the loading control. (D) SiHa cells were treated with VEGF-C (100 ng/mL) for 48 h, in the presence or absence of scrambled siRNA or Moesin siRNA. Cells were stained and analyzed by immunofluorescence. Green, white and yellow arrows indicate lamellipodia, pseudopodia and focal adhesion complexes, respectively. All the experiments were repeated three times with consistent results, and a representative result is shown.