Fig. 4.

RIPK4-mediated BC BIU87 cell EMT and invasion/migration are partly inhibited after silencing of VEGF-A or NF-κB-p65 by specific shRNAs. a BIU87 cells were infected with lentivirus-expressing VEGF-A shRNA-1, shRNA-2, shRNA-3 and shRNA-4, or a control shRNA, VEGF-A protein levels were measured by western blot. b BIU87 cells were infected with lentivirus-expressing NF-κB-p65 shRNA-1, shRNA-2, shRNA-3 and shRNA-4, or a control shRNA, the NF-κB-p65 protein level was measured by western blot. c Western blot showing that after silencing of VEGF-A or NF-κB-p65 in RIPK4-BIU87 cells, the levels of the nuclear NF-κB-p65 and VEGF-A decreased. β-actin was used as a loading control. d Western blot showing that after silencing of VEGF-A or NF-κB-p65 in RIPK4-BIU87 cells, the levels of the mesenchymal markers (vimentin and fibronectin) decreased and the levels of the epithelial markers (E-cadherin and β-catenin) increased. β-actin was used as a loading control. e Wound healing assays showing that the enhanced migratory ability of RIPK4-BIU87 cells was inhibited by silencing of VEGF-A or NF-κB-p65. f The invasive ability of RIPK4-BIU87 cells was inhibited dramatically after shVEGF-A or shNF-κB-p65 treatment in a transwell assay. Data are the means ± SE of three independent experiments. g Representative images of human umbilical vein endothelial cells (HUVECs) cultured with conditioned medium derived from the indicated cells. *P < 0.05 by Student’s t test