Fig. 4

E-cadherin recruits β-catenin away from the Wnt pathway. a Immunofluorescence (IF) for β-catenin (red), E-cadherin (green), and 4′,6-diamidino-2-phenylindole (DAPI) (blue) in triple knockout hepatocellular carcinoma (TKO HCC) and Wnt3a+ L cell (a positive control for nuclear β-catenin). Scale bar = 25 μm. b Immunoprecipitation (IP) for immunoglobulin G (IgG) and β-catenin in TKO HCC cells. The presence of β-catenin, E-cadherin, and α–catenin were detected in the pull-down fraction by immunoblot. c IP for IgG and β-catenin in TKO HCC cells treated with Wnt3A media. The presence of β-catenin and E-cadherin were detected in both unbound and pull-down fraction by immunoblot. Double arrowheads indicate two migrating forms of E-cadherin. d Immunohistochemistry (IHC) for E-cadherin in control liver (CTRL) and TKO HCC. The tumor zone in TKO HCC is delineated by a white dotted line. Scale bar = 100 μm (top), 25 μm (bottom). e Quantitative reverse transcription PCR (RT-qPCR) analysis in control livers (CTRL, n = 4) and TKO HCC (n = 8) for cdh1, snai2, snai1, and twist. f Correlation between E-cadherin vs. β-catenin (left top; p value = 2.509e−12), cdh1 vs. ctnnb1 (left bottom; p value = 4.861e−3), cdh1 vs. E-cadherin (right top; p value = 2.426e−16) and ctnnb1 vs. β-catenin (right bottom; p value = 0.691) expressions in HCC patients at different stages of the disease (CR = correlation ratio; Pearson’s product-moment correlation). g, h Analysis of protein and mRNA expression from The Cancer Genome Atlas (TCGA) patient dataset for g HCC and h other gastro-intestinal cancers (colorectal, stomach, and esophageal). Samples were separated based on E-cadherin protein expression (low vs. high). Expression of glul, tbx3, lgr5, and axin2 were assessed between these two groups to determine the correlation (non-parametric t test) between E-cadherin expression and Wnt target gene expressions in different cancers. Data are represented as mean ± SEM. *p < 0.05 and **p < 0.01. n.s. not significant. See also Supplementary Fig. 4