Figure 3

The activated canonical Wnt signaling of colon cancer cells induces TAM phenotypes of AMSCs in co-culture model. The expression of Wnt signaling molecules and its target genes in HCT116 and AMSCs of co-culture model was detected for examining Wnt activation in these cells. (A, B) The alteration of transcripts of Wnt3a and beta-catenin as well as protein expressions of actived actived β-catenin, phosphorylated (Ser37) β-Catenin and phosphorylated (Tyr279/Tyr216) GSK3β (A), and indicated Wnt target genes of different colon cancer cells or in co-culture models (B) was determined by a qRT-PCR assay. (C) Representative images of confocal fluorescent microscopy of immunofluorescent staining for Wnt3a and beta-catenin of AMSCs cells cultured in indicated conditions. The scale bar represents 20 μm. (D) Immunoblotting assay for indicated Wnt signaling and its target genes in cells cultured in an indicated conditions. GAPDH and Histone H3 in A severed as loading controls for cytosolic and nuclear proteins, respectively. Top panel: immunoblots of indicated proteins; bottom panel: fold changes of interest proteins in top panel quantified by a densitometry analysis. Compared to the control, *p < 0.05; **p < 0.01; ***p < 0.001.