Figure 3.

VASP induces EMT and increases nuclear translocation of β-catenin. (A) Western blot analysis was used to compare expression of epithelial and mesenchymal markers between VASP-transfected cells and shVASP-transfected cells. GAPDH was used as the loading control. (B) Level of β-catenin in nuclear and cytosolic fraction was detected by Western blot analysis. Histone H3 and GAPDH were used as loading controls. (C) LO2 and Hep3B cells were transfected with VASP expression vector and MHCC-97H cells were transfected with VASP shRNA. 72 h after transfection, the expression of cytosolic and nuclear levels of β-catenin (red) in the cells were analyzed by triple IF staining. (D) Immunofluorescence assay was performed to compare the expression patterns of E-cadherin (green) and N-cadherin (red). The nuclei were stained with DAPI in blue color. (E) Immunoblots were performed to detect expression of indicated molecules in Hep3B LV-VASP or Huh7 LV-VASP cells transfected with siRNA targeting Twist1 (Twist1 siRNA) or scrambled siRNA (Control siRNA). (F) Representative pictures of immunochemical staining of serial lung sections for VASP, E-cadherin, N-cadherin, Vimentin, and MMP9 in the Hep3B-LV-control group and the Hep3B-LV-VASP group.