Fig. 3

rNFA47630-induced TNF-α, IL-10, IL-12, and IFN-γ secretion depended on the phosphorylation statuses of MAPK pathways. RAW264.7 cells were stimulated with 2 μg/mL rNFA47630 (with or without PmB) or LPS (with or without PmB) for 30 min (A), or 2 μg/mL protein for 30, 60, or 120 min (B), or 2, 4, or 8 μg/mL protein for 30 min (C), and the phosphorylation statuses of ERK1/2, JNK and p38 were analyzed by western blot. D Cells were incubated with 2 μg/mL rNFA47630 (with or without PmB) or LPS (with or without PmB) for 6, 18, and 24 h, and the levels of TNF-α, IL-10, IL-12, and IFN-γ in the supernatants were measured by ELISA (n = 6 in each group). E Cells were pretreated for 1 h with inhibitors of PD 98059, SP 600125 or SB 203580 prior to rNFA47630 protein exposure, and cytokine levels were measured by ELISA (n = 6 in each group). Data are expressed as the means ± SDs for three independent experiments. p < 0.001 when compared with the control group (D) or rNFA47630 group (E)