Fig. 4.

TLR2 and TLR4 mediated IL-6, IL-23 and IL-17 production in neutrophils. (a–d) Neutrophils were isolated from total bone marrow cells of WT, Tlr2^−/− and Tlr4^−/− mice, and then infected with MTB H37Rv. (a) Immunoblot analysis of phosphorylated p-p65, p-AKT, AKT, p-p38, p38, p-ERK, ERK, p-JNK, JNK and GAPDH at 30 min after infection. (b) IL-6 and IL-23 concentrations were quantified by ELISA in supernatants at 12 h. (c) Il7a expression was evaluated by real-time PCR at 18 h and IL-17 concentrations in supernatants were quantified at 24 h. All gene quantities were normalized to the expression of the reference gene β-actin. (d) Expression of intracellular IL-17 at 24 h. (e) Neutrophils were isolated from total bone marrow cells of WT, Tlr2^−/− and Tlr4^−/− mice, and then stimulated with Pam3CSK4, LPS or mixture of Pam3CSK4 and LPS (Mix). IL-6 and IL-23 concentrations were quantified by ELISA in supernatants at 12 h. (f) Il7a expression was evaluated by PCR in WT neutrophils after stimulated with Pam3CSK4, LPS or mixture of Pam3CSK4 and LPS. Data shown are the mean ± SEM. *P < 0.05; **P < 0.01. Data are representative of three independent experiments with similar results.