Fig. 7. Suppressive effect of Wnt-C59 on the interaction between β-catenin and NF-κB along with the activities of Nuclear factor-kappa B (NF-κB) and its responsive promoter in BEAS-2B human bronchial epithelial cells.

Cells were treated with 0, 30, or 50 μM of Wnt-C59, followed by lipopolysaccharide (LPS) stimulation at 0.1 μg/ml for 2 h. (A, C) The protein-protein interaction between β-catenin and NF-κB p65 was analyzed by co-immunoprecipitation assays followed by Western blotting. Original uncut Western blot images were shown in Supplementary Data 2. Input samples of co-immunoprecipitation assays are shown in Supplementary Data 3. (B, D) The amount of co-immunoprecipitated β-catenin or NF-κB p65 was measured by ImageJ. (E) NF-κB activity for its target DNA binding was measured by ELISA. (F) Transcriptional activity of NF-κB-responsive promoter was measured by luciferase reporter assay. *p < 0.05, **p < 0.01, ***p < 0.001 compared with cells stimulated with LPS with 0 μM of Wnt-C59. ^##p < 0.01, ^###p < 0.001 compared with unstimulated cells. Experiments were conducted in triplicate. Data are shown as mean ± standard deviation, and statistical significance was measured by unpaired t-test. IP, immunoprecipitation; WB, Western blot.