Fig. 2

WNT4 induces the expression of β-catenin. a and b Cell migration and invasion capabilities of colorectal cancer (CRC) cells pretreated with WNT4 (100 ng/mL) for 24 h. Scale bar, 50 μm. c TOP-flash plasmid or FOP-flash (2 mg) transfected into the CRC cells for 48 h. Cells were treated with WNT4 (100 ng/mL) for 6 h before analysis. d LoVo and HCT 116 CRC cells treated with WNT4 (100 ng/mL); and the expression of β-catenin, AXIN2, E-cadherin, ZO-1, and nuclear β-catenin and AXIN2 were determined by western blot analysis. e Higher expression and nuclear translocalization of β-catenin and AXIN2 were shown in immunofluorescence after LoVo cells treated with WNT4 (100 ng/mL) for 6 h. Scale bar, 10 μm. f and g TOP/FOP-Flash reporter assay and western blots were used to detect whether the WNT pathway was activated. CRC cells were treated with ICG-001 (25 μM) for 24 h and WNT4 (100 ng/mL) for 6 h. Two-tailed Student’s t test and ANOVA were used for statistical analyses. All experiments were performed in triplicate. Measurement data were presented as the mean ± SD. *P < 0.05; NS, no significance