The JNK and STAT3 signaling pathways play an important role in IL-22-mediated inflammatory cytokine production by BV2 and HT22 cells, respectively. (A) RT-PCR analyses of TNF-α in cells that were pretreated with DMSO (vehicle control), SP600125 (20 μM), and S3I-201 (50 μM) for 1 h prior to treatment with IL-22 (20 ng/mL) for 12 h. (B) ELISA-based analysis of TNF-α in the supernatants of cells that were pretreated with DMSO (vehicle control), SP600125 (20 μM), or and S3I-201 (50 μM) for 1 h prior to treatment with IL-22 (20 ng/mL) for 48 h. ***
p < 0.001. (C) Immunoblot analyses of c-Jun and phosphorylated c-Jun in BV2 cells that were pretreated with SP600125 (20 μM) and then treated with IL-22 (20 ng/mL) for 0, 5, 10, 20, 30, or 60 min. Relative intensity was analyzed by ImageJ software. All results were representative of at least three independent experiments. Values were presented as the mean ± SD. Significance (p-value) was determined by t-test, ***
p < 0.001. (D) Immunoblot analyses of STAT3 and phosphorylated STAT3 in HT22 cells that were pretreated with S3I-201 (50 μM) and then treated with IL-22 (20 ng/mL) for 0, 5, 10, 20, 30, or 60 min. (A,C,D) Results are representative of three independent experiments. Relative intensity was analyzed by ImageJ software. All results were representative of at least three independent experiments. Values were presented as the mean ± SD. Significance (p-value) was determined by t-test, ***
p < 0.001.