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. 2017 Aug 9;23:88–99. doi: 10.1016/j.ebiom.2017.08.001

Fig. 5.

Fig. 5

Expression of IL-6, IL-23 and IL-17 in human peripheral blood neutrophils. (a–e) Neutrophils were isolated from human peripheral blood, and then infected with MTB H37Rv. The uninfected neutrophils were as control group. (a) Il6 and Il23mRNA expression for indicated time points determined by real-time PCR. All gene quantities were normalized to the expression of the reference gene gapdh. (b) Intracellular IL-17 identified by Flow cytometry. (c) Il17a expression plus no antibody (-) or neutralizing antibody (Ab) to IL-1β, TGF-β, IL-6 or IL-23. GAPDH (housekeeping gene) serves as a loading control throughout. (d–e) IL-6, IL-23 (d) and IL-17 (e) concentrations in supernatants were quantified by ELISA after treated with NF-κB-inhibitor JSH-23 (20 μM), PI3K- inhibitor LY294002 (30 μM), MEK1/2-inhibitor U0126 (40 μM), p38 inhibitor-SB203580 (10 μM) or JNK-inhibitor SP600125 (20 μM). (f–g) Neutrophils were isolated from human peripheral blood, and then stimulated with Pam3CSK4, LPS or mixture of Pam3CSK4 and LPS. The unstimulated human neutrophils were as control group (Ctrl). (f) IL-6 and IL-23 concentrations in supernatants were quantified. (g) Intracellular IL-17 identified by Flow cytometry. Data shown in are the mean ± SEM. *P < 0.05; **P < 0.01. Data are representative of three independent experiments with similar results.