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. 2016 Aug 31;7(39):64030–64042. doi: 10.18632/oncotarget.11747

Figure 5. CFTR interacts with β-catenin and prevents its degradation in Caco-2 cells and mouse small intestine.

Figure 5

(A) Fluorescent immunostaining of CFTR (1:50, ACL-006, Alomone Labs) and β-catenin (1:200, CST2677, Cell Signaling) in Caco-2 cells shows co-localization of CFTR and β-catenin. Green color: CFTR; Red color: β-catenin; Blue color: To-pro3. Scal bar: 10 μm. (B) Co-immunoprecipitation study on Caco-2 cells. Left panel: both CFTR and NF-κB p65 immunoprecipitate with anti-β-catenin antibody; middle panel: β-catenin and NF-κB p65 immunoprecipitate with anti-CFTR antibody; right panel: CFTR and β-catenin immunoprecipitate with anti NF-κB p65 antibody. The blots shown are representative image from three independent experiments. (C) Proteasome inhibitor MG132 (10 μM) prevents CFTR knock down-induced β-catenin decrease in Caco-2 cells. Quantification data is acquired from three independent experiments. (D) Immunoprecipitation study in the intestine of wild type and ΔF508cftr−/− mice. In wild type mouse intestine, both CFTR and NF-κB p65 are pulled down by anti-β-catenin antibody. In ΔF508cftr−/− mouse intestine, ΔF508 CFTR protein is not co-immunoprecipitated with anti-β-catenin antibody, and the interaction between NF-κB p65 and β-catenin is significantly reduced (**p < 0.01, One-way ANOVA).