Ab109404

For immunofluorescence staining of phosphor‐TAK1, 2 μm frozen colon sections were used. Sections were stained with rabbit monoclonal anti‐TAK1 (ab109404; Abcam) followed by goat anti‐rabbit‐Alexa Fluor‐488 (ab150077; Abcam). For the nuclear counter staining, sections were stained with DAPI (Life Technologies).

The HASMCs and rat tissues were lysed with RIPA buffer containing complete protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific). Protein concentrations were measured using a BCA protein assay kit (Interchim, Montluçon, France). The samples were separated by 10% or 12% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and the resulting protein bands were transferred onto PVDF membranes. After being blocked, the membranes were incubated with the following primary antibodies overnight at 4 °C: NF-κB antibodies (1:1000, ab32536, Abcam), TNF-α antibodies (1:1000, ab6671, Abcam), FXR antibodies (1:500, sc-13063, Santa Cruz Biotechnology), TAK1 antibodies (1:1000, ab109526, Abcam), p-TAK1 antibodies (1:1000, ab109404, Abcam), TAB1 antibodies (1:1000, ab227210, Abcam), and Phospho-IκBα antibodies (Ser32/36) (1:1000, #9246, Cell Signaling Technology). Then, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies at room temperature for 2 hours. The protein bands were detected using an ECL Substrate (Thermo Fisher Scientific), and the quantification of the average densities of the protein bands was performed using the Image-Pro Plus 4.5 software.