FIGURE 3.

The activation of Wnt signaling pathway in tumors arising from Villin‐Cre; Smad4^{F / F} ; Trp53^{F / F} mice. (A) The enrichment of Wnt signaling pathway using GSEA in Villin‐Cre; Smad4^{F / F} ; Trp53^{F / F} intestinal adenocarcinomas (n = 2) compared with Villin‐Cre‐negative normal intestinal mucosa (n = 2) based on mouse gene expression microarray analysis. (B) Representative IHC images for c‐myc and cyclin D1 in primary Villin‐Cre; Smad4^{F / F} ; Trp53^{F / F} intestinal cancer tissues. (C) Representative IHC images for β‐catenin in neoplastic lesions from Villin‐Cre; Smad4^{F / F} ; Trp53^{F / F} mice. (D) IHC grading of nuclear β‐catenin expression in neoplastic lesions from Villin‐Cre; Smad4^{F / F} ; Trp53^{F / F} mice (n = 3). (E) TOPflash reporter activities in primarily cultured cancer cell lines (primary #1 and primary #2) established from Villin‐Cre; Smad4^{F / F} ; Trp53^{F / F} intestinal adenocarcinomas to measure Wnt pathway activities after the restoration of Smad4 and/or p53 using lentiviral system. Right, western blot analysis to confirm the restoration of Smad4 and/or p53 in primary #2 cells. GSEA, gene set enrichment analysis; IHC, immunohistochemistry. *p < 0.05