Figure 6
Involvement of C-Jun N-terminal kinase (JNK) in the regulation of mPGES-1 and COX-2. (A) Gingival fibroblasts were treated with TNFα (20 ng/ml) with or without the JNK inhibitor SP600125 (SP, 10 μM) for 24 h. Expression of mPGES-1 and COX-2 was measured by flow cytometry using specific antibodies. (B-C) Gingival fibroblasts were treated with TNFα (20 ng/ml) with or without SP (10 μM) for 1, 3, 6 and 24 h (B) or 10 minutes (C). Cells were lysed and total protein was analyzed for phosphorylated JNK (p-JNK) and expressed as relative to control cells at 1 h (B) or at start of incubation (C). Data is presented as mean ± s.d. Asterisks (*) indicate a significant difference (p < 0.05) between TNFα-stimulated cells and control cells at each time point, and hash symbols (#) indicate a significant difference (p < 0.05) between TNFα-stimulated cells and cells treated with TNFα in combination with SP at each time point. (D) Gingival fibroblasts were cultured with the indicated doses of SP in the absence or presence of TNFα (20 ng/ml) for 24 h. Levels of PGE2 in the culture media were measured by EIA using Luminex technology. Data is presented as mean ± s.d. Asterisks (*) indicate a significant difference compared to control cells not treated with TNFα or SP, and hash symbols (#) indicate a significant difference compared to cells treated with TNFα only (p < 0.05). The results are representative for all three cell lines and all analyses were performed in triplicates.