Figure 5
Overexpressing suprabasin activates the Wnt/β-catenin signaling pathway.
(a) GSEA analysis showing the correlation between suprabasin expression and Wnt/β-catenin signaling in the published breast cancer dataset (TCGA database). (b) Suprabasin activates the Wnt/β-catenin signaling activity, measured by luciferase assay of TCF/LEF transcriptional activity in the indicated cells. (c) Nuclear expression of β-catenin in indicated ESCC cell lines, determined by Western blotting using p84 as a nuclear marker. (d) Real-time PCR analysis of Wnt/β-catenin signaling pathway downstream genes expression, including AXIN2, CTLA4, CCND1, FGFs, FRA1, ITF2, MMP7, JUN, LEF1, MYC and TCF1 mRNAs in the indicated ESCC cells. (e) Western blotting of phosphorylated β-catenin (phospho S33 ± S37), β-catenin, Cyclin D1 and c-Myc in ESCC cells. GAPDH was used as the loading control. (f,g) Quantification of colonies determined by colony formation (f) and anchorage-independent growth assays (g), after cells treated with IWR-1-endo (Wnt/β-catenin response inhibitor, 10μM) or vehicle control. (h) Representative images of the tumors from all mice in each group. Bars represent the mean ± SD of three independent experiments. *P < 0.05.
