Fig. 2

CircANKS1B enhances the migratory, invasive and metastatic capabilities of breast caner cells by inducing EMT. a qRT-PCR for the expression of circANKS1B in breast epithelial cell and breast cancer cells. b Wound healing assays for MCF-7 cells with circANKS1B overexpression and MDA-MB-231 cells with circANKS1B knockdown. Representative images are shown at 0 and 24 h after wound creation. Scale bar = 20 μm. c Transwell migration and matrigel invasion assays for MCF-7 cells with circANKS1B overexpression and MDA-MB-231 cells with circANKS1B knockdown. Scale bar = 20 μm. d The images of lung of nude mice in five treatment groups. Black arrowheads indicate the lung metastatic nodules stained by hematoxylin and eosin. Scale bar = 20 μm. e Representative phase-contrast images of circANKS1B-overexpressing MCF-7 cells and circANKS1B knockdown MDA-MB-231 cells. Scale bar = 20 μm. f-g Immunoblot analysis of E-cadherin, Vimentin and Fibronectin protein expression in circANKS1B-overexpressing MCF-7/T47D cells and circANKS1B silencing MDA-MB-231/BT549 cells. GAPDH was used as a loading control. Data were represented as means ± S.D. of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001