Fig. 7

The identified β-catenin residues targeted for phosphorylation by p38α are crucial for β-catenin transcriptional activity. A Co-immunoprecipitation of p38α with FLAG-β-catenin-WT or FLAG-β-catenin-S111A or FLAG-β-catenin-T112A in HCT-116 cells. Input corresponds to 10% of the lysate. Anti-IgGs were used as negative controls. B Chromatin immunoprecipitation assay in HCT-116 cells. Cells overexpressing FLAG-β-catenin-WT or FLAG-β-catenin-S111A or FLAG-β-catenin-T112A were treated or not with ralimetinib (10 μM) for 24 h. Chromatin was pulled down with anti-FLAG antibodies. Anti-IgGs were used as negative controls. *P < 0.05 vs. FLAG-β-catenin-WT; ^#P < 0.05 vs. DMSO. C Quantification results of the ddPCR assay (copies/µL) of β-catenin and c-Myc mRNA expression, as processed by QuantaSoft. HCT-116 CRC cells silenced by genetic ablation for endogenous β-catenin and exogenously expressing β-catenin-WT, β-catenin-S111A, β-catenin-T112A were treated or not with the p38α inhibitor ralimetinib (10 μM) for 24 h. The error bars represent the maximum and minimum Poisson distribution for the 95% confidence interval generated by QuantaSoft. Results are representative of at least three independent experiments