Fig. 3

Fluda inhibited iNOS, NF-κB, and ERK signaling. A The mRNA expression of Nos2 was detected in BAL cell lysates using real-time PCR. B The protein expression of iNOS was detected using western blotting (left). Densitometric ratio of iNOS against β-actin (right). C Release of NO was determined by measuring the amount of nitrite in BALF using Griess reagent. D iNOS protein expression in lung tissue was detected using western blotting (upper). Densitometry ratio of iNOS in tissue lysates against β-actin (bottom). E The expression of phosphorylated NF-κB (Ser536), JNK1/2, ERK1/2, and p38 was detected. Total NF-κB, total JNK1/2, total ERK1/2, and total p38 were used as loading controls. The proteins were isolated from mice lung tissue. Densitometric ratios of pNF-κB, pERK1/2, pJNK1/2, and p-p38 in tissue lysates against loading controls. The data are presented as mean ± SD (n = 3 per group for A-C and n = 4 per group for D and E). **p < 0.01, and ***p < 0.001 compared with the control group. †p < 0.05, ††p < 0.01, and †††p < 0.001 compared with the LPS group. ns: not significant