Fig. 4

Elevation of EHMT2 promotes Wnt/β-catenin pathway activity. A The subcellular localization of β-catenin was determined by immunofluorescence staining. Scale bar, 50 μm. n = 6 (mean ± SD); ***p < 0.001 by student's t-test. B TOP/FOP luciferase showed Wnt–β-catenin signaling activity in Hep3B and Huh1 cells. Cells (wild type and EHMT2^−/− single clones in 2A) were treated with Wnt3a conditional medium (CM) at the indicated concentration. n = 6 (mean ± SD); *p < 0.05 and ***p < 0.001 by one-way ANOVA. C TOP/FOP luciferase showed Wnt–β-catenin signaling activity in Hep3B and Huh1 cells. Cells in 3B were pre-treated with UNC0642 (5.0 μM for 8 h) and then subjected to Wnt3a CM at the indicated concentration. n = 6 (mean ± SD); n.s, no significance, *p < 0.05 **p < 0.01 and ***p < 0.001 by student's t-test. D RT-qPCR showed Wnt–β-catenin signaling target gene expression in Hep3B cells. Cells in 3B were treated with 25% Wnt3a conditional medium. n = 6 (mean ± SD); n.s, no significance, *p < 0.05, **p < 0.01 and ***p < 0.001 by student's t-test. E The nuclear β-catenin protein level in cells was determined by Western blot. Cells and treatment were the same as 4D, and the nucleus of harvested cells was isolated. n = 6 (mean ± SD); **p < 0.01 and ***p < 0.001 by student's t-test