FIG 3 .

NhhA-triggered Mo differentiation is mediated by IL-10 and TNF-α. (A) Mo were stimulated with 50 nM NhhA alone (control) or in the presence of blocking antibodies (Abs) (5 µg/ml) to the indicated human TLRs. Some cells were cotreated with 500 nM Celastrol (an NF-κB inhibitor). RNA was extracted after 3 h, and the mRNA levels of the indicated cytokines were quantified by real-time PCR (qPCR). The data were normalized to the reference gene rpl37A and are presented as fold change relative to that of Mo without NhhA stimulation, which was set to 1. Data shown are the mean ± standard deviation from three independent experiments; *, P < 0.05 compared with the control using ANOVA followed by the Bonferroni post hoc test. (B) Mo were stimulated with 50 nM NhhA alone (control) or in the presence of the blocking agent for the indicated cytokines as described in Materials and Methods. The differentiation state was assessed by the flow cytometric analysis of CD14 expression and SSC parameters (shown as median fluorescence intensity [MFI]). Error bars indicate medians with interquartile ranges of three independent experiments. Data are presented as fold change relative to control; *, P < 0.05 compared with the control using ANOVA followed by the Bonferroni post hoc test.