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 4. P22077 inhibits the activation of NF-κB and MAPKs signaling pathways in LPS-stimulated Raw264.7 cells and mouse peritoneal macrophages. (A) Raw264.7 cells were pretreated with DMSO and P22077 (7.5 μM) for 2 h, and then stimulated with LPS (100 ng/ml) for the indicated times and analyzed the indicated proteins by immunoblot. (B) Mouse peritoneal macrophages were pretreated with DMSO and P22077 (7.5 μM) for 2 h, and then stimulated with LPS (100 ng/ml) for the indicated times and analyzed the indicated proteins by immunoblot. (C) RAW 264.7 cells were transfected with NF-κB luciferase reporter plasmid and Renilla luciferase construct phRL-TK plasmid for 24 h and pretreated with the P22077 (7.5 μM) for 2 h before stimulation with 100ng/ml LPS for 4 h and luciferase activity was measured. Similar results were obtained from three independent experiments and data were presented as mean ± SD of one representative experiment. **P<0.01 vs LPS-stimulated group.