Research Paper Volume 12, Issue 11 pp 10969—10982

P22077 inhibits LPS-induced inflammatory response by promoting K48-linked ubiquitination and degradation of TRAF6

Figure 2. P22077 suppresses LPS-induced inflammatory response in Raw264.7 cells. (A, B) Raw264.7 cells were pretreated with DMSO or P22077 (2.5 μM, 5.0 μM and 7.5 μM) for 2 h, then stimulated with LPS (100 ng/ml) for another 4 h. The mRNA expressions of TNF-α, IL-1β, IL-6 (A), COX2 and iNOS (B) were analyzed by Q-PCR. (C, D) Raw264.7 cells were pretreated with DMSO or P22077 (2.5 μM, 5.0 μM and 7.5 μM) for 2 h, and then stimulated with LPS (100 ng/ml) for 10 h. Nitric oxide (NO) concentration in culture supernatant was measured by Griess assay (C). TNF-α and IL-6 in the supernatants were measured by ELISA, pro-IL-1β was detected by immunoblot (D). Similar results were obtained from three independent experiments and data were presented as mean ± SD of one representative experiment. *P<0.05, **P<0.01, ***P<0.001 vs LPS-stimulated DMSO group.