Figure 5

CO inhibits LPS-induced COX-2 protein expression and PGE ₂ synthesis in bEnd.3 cells. A: Cells were pretreated with CO-RM2 (CO releasing molecule) for 2 h and then incubated with LPS (100 mg/ml) for 24 h. The expression of COX-2 was determined by western blot. *P < 0.05; ^#P < 0.01, as compared with the cells incubated with LPS alone. B: Cells were pretreated with CO-RM2 (50 mM, 2 h) and then incubated with LPS (100 mg/ml) for 24 h. The media were used to determine the levels of PGE₂. *P < 0.05; ^#P < 0.01, as compared within groups. C: Cells were pretreated with CO-RM2 (CO releasing molecule) for 2 h and then incubated with LPS (100 mg/ml) for the indicated time intervals. Phosphorylation of NF-κB p65 was determined by western blot. *P < 0.05; ^#P < 0.01, as compared within groups. NS: not significant. D: Cells were treated with or without CO-RM2 (50 μM, 2 h) followed by LPS treatment (100 mg/ml) for the indicated time intervals. Cell lysates were immunoprecipitated with an anti-TLR4 antibody. The immunoprecipiates were analyzed by western blot using anti-MyD88 or anti-TLR4 (input) antibody. Data are summarized and expressed as mean ± SEM for four individual experiments. *P < 0.05 as compared within groups.