Figure 4

The phosphorylation of STAT3 was crucial for the IL-22-mediated production of TRIM25. A DEFs cells were stimulated with different amounts of IL-22 and WP1066 and were harvested 24 h post-treatment. Then, 30 mg of protein lysates were analyzed for P-STAT3 (S727) and total STAT3 via Western blot analysis (anti-pSTAT3 1:3000, anti-STAT3 1:3000, anti-β-actin 1:4000, HRP-conjugated goat anti-mouse antibody 1:3000). B The inhibition of P-STAT3 suppressed TRIM25 production. DEFs were treated with 5 µM WP1066 (or PBS in the control group) and IL-22 at 50 ng/mL for 24 h; TRIM25 expression was then measured via qRT-PCR and Western blotting. The results are expressed as the mean ± SEM of three independent experiments. *P < 0.05, ***P < 0.001.