Fig. 7
From: Inhibition of transmembrane TNF-α shedding by a specific antibody protects against septic shock
tmTNF-α Ab inhibits the TRIF-dependent TLR4 signaling pathway. THP-1-derived or Raw264.7 macrophages were stimulated with 100 ng/ml LPS, combined with 2 μg/ml tmTNF-α mAb or pAb, respectively. Isotype antibody IgG or normal serum IgG served as a control. a, d Representative western blot analysis of three independent experiments for IRF3 phosphorylation in THP-1-derived (1 h after stimulation) or Raw264.7 macrophages (45 min after stimulation). Relative levels of IFN-β mRNA were assessed by real-time PCR at 4 h (b, e), and IFN-β release was detected by ELISA at 10 h (c, f) after stimulation. All quantitative data are presented as means ± SEM of at least three independent experiments. Mice were intraperitoneally injected with 600 μg tmTNF-α pAb or normal serum IgG immediately after the CLP operation (n = 6 each group). Western blot analysis of IRF3 phosphorylation (g) and relative levels of IFN-β mRNA in the liver (h) and plasma levels of IFN-β (i) at 24 h after the CLP operation. *p < 0.05, **p < 0.01, ***p < 0.001
