Fig. 3

Uncoupling p38α cytoplasmic and nuclear functions in the Wnt pathway. A, H Immunoblotting analysis of p38α and β-catenin cellular localization in HT-29 (A) and HCT-116 (H) CRC cells under serum starvation (24 h) and upon activation of the Wnt pathway mediated by serum supplementation (A, H) or addition of Wnt3a (50 ng/mL) and the GSK3β inhibitor TWS-119 (10 μM) for 4 h (A). Subsequently, cells were treated or not with the p38α inhibitor ralimetinib (10 μM) for 24 h (A, H). B–F Densitometric analysis of the indicated protein levels against the loading control in the different culture conditions used in this study. Statistical analysis was performed using Student’s t-test: *P < 0.05 vs. no serum (B, C) or vs. no ralimetinib (D–F). G RTqPCR analysis of β-catenin target gene expression in HT-29 cells treated as in A. Statistical analysis was performed using Student’s t-test: *P < 0.05 vs. no serum; ^#P < 0.05 vs. no ralimetinib. Lamin B1: nuclear loading control; PDI: cytoplasmic loading control. N = Nucleus, C = Cytoplasm. Results are representative of at least three independent experiments