Fig. 2.

Fig. 2. The iSCAP-KD cells are significantly less responsive to Wnt3A-activated canonical signaling pathway. (A) iSCAP-KD cells exhibit significantly lower β-catenin/Tcf reporter activity upon Wnt3A stimulation. Subconfluent iSCAP-KD and iSCAP-Ctrl cells were transfected with TOP-Luc reporter plasmid and infected with AdWnt3A or AdGFP. At 48h post transfection/infection, cells were lysed for luciferase assays. Easy conditions were done in triplicate. (B).Wnt3A-induced expression of Wnt/β-catenin target genes was significantly decreased in iSCAP cells. Subconfluent iSCAP-KD and iSCAP-Ctrl cells were infected with AdWnt3A or AdGFP for 36h. Total RNA was isolated and subjected to reverse transcription. The resultant cDNAs were used as templates for qPCR analysis using primers specific for mouse Axin2 and c-Myc transcripts. All samples were normalized by GAPDH levels. Each assay condition was done in triplicate.