Fig. 3

ATF2 facilitates RCC cell migration and invasion in vitro. a Left: representative images of the wound-healing assay of ATF2 knockdown and control RCC cells photographed at 0, 6 and 12 h after scratching. Scale bar = 200 μm. Right: the relative migration rate was calculated by dividing the change in the distance between the scratch edges by the initial distance. b & c Transwell assays were performed to evaluate cell migration following ATF2 knockdown (b), ATF2 overexpression (c) and control RCC cells. The statistical graph indicates the means ± SEM of the number of cells from 8 random high power fields (magnification, ×200) counted from three independent experiments. Scale bar = 2 mm. d & e Transwell assays were performed to evaluate the invasion of ATF2 knockdown (d), ATF2 overexpression (e) and control RCC cells. The statistical graph indicates the means ± SEM of the number of cells from 8 random high power fields (magnification, ×200) counted from three independent experiments. Scale bar = 2 mm. f & g Western blotting analysis of Snail, Vimentin and E-Cadherin in ATF2 knockdown (f), ATF2 overexpression (g) and control 786-O cells. β-Actin was used as an internal standard. h ChIP assay analysis of the enrichment of ATF2 at the proximal promoter region of Snail and Vimentin in the context of ATF2 overexpression. The enrichment of ATF2 on Snail and Vimentin promoter relative to input in 786-O cells (upper). The gel electrophoresis of PCR products from ChIP assay (lower). Results are presented as mean ± SEM from three independent experiments. *p < 0.05, **p <0.01, and ***p < 0.001