Fig. 5

NCSTN promotes nuclear translocation of β-catenin. a Immunofluorescence assays showed NCSTN increased nuclear translocation of β-catenin. Scale bars, 100 μm. b Subcellular fractionation assays showed the expression of β-catenin in the cytosol and nuclear of indicated cells. Loading control was assessed by β-tublin and Histone H3. c TOP/FOP Flash assays showed the transcriptional activity of β-catenin in the indicated HCCLM3 and Hep3B cells. d The expression levels of β-catenin target protein c-myc, cyclin D1 and mmp-7 in the indicated cells. e Representative immunohistochemical staining images of NCSTN and β-catenin in HCC tissues (left), correlation between NCSTN expression and nuclear β-catenin expression (right). Scale bars, 100 μm (left), 25 μm (right). f Schematic outlines of the predicted binding sites of β-catenin on Zeb1 gene promoter region. g ChIP assays of the enrichment of β-catenin on BSs in the promoter region of Zeb1 relative to IgG. h ChIP assays showed relative enrichment of β-catenin on BS1 in the Zeb1promoter region in the indicated cells. i The expression levels of EMT-related protein in the indicated Hep3B cells co-transfected with or without siβ-catenin. j CCK8 assays showed that NCSTN-mediated cell growth could be rescued in Hep3B cells co-transfected with siβ-catenin. k The migration and invasion capacity was rescued in the indicated Hep3B cells co-transfected with siβ-catenin. BS, binding sites. *p < 0.05, **p < 0.01, ***p < 0.001