Fig. 4

NCSTN induces EMT in HCC cells via activation of Zeb1. a The mRNA expression levels of EMT markers CDH1, CDH2, VIM, ZEB1, SNAIL1, SNAIL2, FOXC1, FOXC2 and TWIST1 in the indicated HCC cells. b Luciferase reporter assays showed relative luciferase activity of Zeb1 promoter in indicated cells. c The expression levels of four EMT-related proteins E-cadherin, Zeb1, Vimentin and N-cadherin in the indicated cells. Loading control was assessed by β-actin. d Immunofluorescence assays showed the expression of NCSTN, E-cadherin, Vimentin and Zeb1 in the indicated cells. Scale bars, 100 μm. e The expression levels of EMT-related markers E-cadherin, Vimentin and N-cadherin in the indicated cells co-transfected with siZeb1 or negative control. Loading control was assessed by β-actin. f The migration and invasion capacity was examined in the indicated HCC cells co-transfected with siZeb1 or negative control. g Wound healing assays showed the migration capacity of indicated HCC cells. h Representative images of EMT-related markers E-cadherin, Vimentin and N-cadherin in HCC tissues by immunohistochemical staining. Scale bar, 100 μm. i Correlation analyses of protein expression levels between NCSTN and E-cadherin, Vimentin or nuclear Zeb1 in HCC tissues. *p < 0.05, **p < 0.01, ***p < 0.001