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. 2017 Jan 20;7:40984. doi: 10.1038/srep40984

Figure 5. ESAT-6 stimulates STAT3 phosphorylation and IL-6 production by macrophages independent of TLR2 or autocrine effects of IL-6 and IL-10.

Figure 5

BMDMs from three wild-type or TLR2−/− mice were incubated with ESAT-6 for different time points and the phosphorylation of STAT3 was determined by Western blotting followed by blotting for total STAT3 in the same membrane after stripping (A). BMDMs from 5 wild-type (filled bars) or 6 TLR2−/− (open bars) mice were incubated with medium alone or indicated concentrations of ESAT-6, LPS or Pam3CSK4 and IL-6 levels were determined 24 h later (B). Data are expressed as means ± SEM. BMDMs from wild-type mice were incubated with either anti-IL-6 or anti-IL-6R for one hour prior to addition of ESAT-6 at 1 ug/ml for different time points. The phosphorylation of STAT3 and total STAT3 (C) was determine as in (A) and IL-6 production (D) was determined as in (B). BMDMs were incubated with anti-IL-10 at 5 μg/ml for one hour prior to stimulation with ESAT-6 or infection with H37Rv. Activation of STAT3 (E) and production of IL-6 (F) were determined as in (A and B), respectively. One representative result of three independent experiments is shown for (A,C and E). The data are expressed as means ± SEM for (B,D and F).