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. 2015 Aug 18;17(7):598–611. doi: 10.1016/j.neo.2015.07.006

Figure 3.

Figure 3

Wnt/β-catenin signaling in esophageal cell lines: (A) qRT-PCR analysis of AXIN2, Dkk1, Cyclin D1, and c-MYC mRNA expression in two squamous esophageal cell lines (EPC1/EPC2), one BE metaplasia cell line (CP-A), and esophageal adenocarcinoma cell line (OE33) in resting conditions. (B) Luciferase assay in all cell lines in resting conditions. TOP-flash, and not FOP-flash, is responsive to co-activation of TCF/LEF by β-catenin. The colorectal cancer cell line Caco2 was used as positive control. Relative mean gene expression to that of EPC1 cells is presented. (C) Western blot analysis of ABC and Dkk1 protein expression in all cell lines at resting conditions. β-Actin serves as a loading control. (D) Immunofluorescence microscopy for colocalization of total β-catenin (Ab against C-terminus) and ABC protein in all cell lines at resting conditions. ABC was predominantly nuclear, whereas total β-catenin was localized in the membrane and adherent junctions. Representative tissue staining of at least three different tissues is presented [comparisons between cell lines for Cyclin D1#, c-MYC&, and Dkk1@; #, &, @P < .05; **P < .01;***P < .001; t test).