Fig. 2

Claudin5 knockdown promotes proliferation, invasion and migration of ESCC cells in vitro and in vivo. A, B Claudin5 expression levels were assessed in TE1 cells transfected with Claudin5 siRNA (A), and in KYSE150 cells transfectsed with Claudin5 (B) using qRT-PCR and western blotting. GAPDH served as the loading control. C, D The effects of Claudin5 knockdown (C) or overexpression (D) on the ESCC cells proliferation were evaluated by CCK-8 assay. E, F The effects of Claudin5 knockdown (E) or overexpression (F) on ESCC cells invasion were evaluated by transwell invasion assay (magnification, × 40). G, H The effects of Claudin5 knockdown (G) or overexpression (H) on ESCC cells migration were evaluated by wound healing assay (magnification, × 200). I Representative data of tumors in nude mice bearing TE1 cells. J Tumor weight of xenograft mouse tumors. K Tumor volumes of xenograft mouse tumors. L, M Claudin5 knockdown resulted in increased lung metastasis, shown by the elevated number of lung nodules. The statistical difference was assessed with one-way ANOVA followed by Dunnett tests in A, C, E, G, J, K and M; and the two-tailed unpaired Student t test in B, D, F and H. Error bars show the SD from three independent experiments. **p < 0.01, ***p < 0.001, ****p < 0.0001