Fig. 3. M1 prevents LPS-induced TNFα production in peritoneal macrophages via inhibiting TLR4/NFκB signal activation.

Murine peritoneal macrophages were treated with different concentration of M1 (1, 10, 100 μM) 1 h before the addition of 10 μg/ml LPS for 6 h. The protein level a and mRNA expression b of TNFα were assayed by western blot and QPCR, respectively (n = 3). c Then the relative expression of phosphor-NFκB, NFκB, phosphor-IκB, and IκB were quantified as well (n = 3). d NFκB activation were also determined by a luciferase reporter gene transfected in RAW264.7 cells (n = 3). e TLR4 expression on the surfaces of murine macrophages were detected by flow cytometry (n = 3). f Hepatic expression of TLR4, phosphor-IκB, and IκB were analyzed by western blot (n = 5). Each value is mean ± S.E.M. *P < 0.05, **P < 0.01, ***P < 0.001