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. 2019 Mar 12;9:4246. doi: 10.1038/s41598-019-38982-0

Figure 2.

Figure 2

Rv3463 induces bone marrow-derived macrophages (BMDMs) activation. BMDMs were stimulated with Rv3463 (1, 3 or 5 μg/ml), lipopolysaccharide (LPS, 100 ng/ml), Ag85 (5 μg/ml), or ESAT-6 (2 μg/ml) for 24 h. (A) The cytokines from the culture supernatants were measured by ELISA. All data are expressed as mean ± SD (n = 3). (C) BMDMs stimulated with each antigen were also analyzed for the expression of surface markers by two-color flow cytometry. The cells were gated to exclude F4/80+ cells. BMDMs were stained with anti-CD80, anti-CD86, anti-MHC class I or anti-MHC class II antibodies. The histograms are representative of five experiments. Bar graphs show the percentage (mean ± SD of five experiments) for each surface molecule on F4/80+ cells. (B) BMDMs were incubated with LPS (100 ng/ml) or Rv3463 (5 μg/ml) with or without pretreatment of polymyxin B (PMB) for 1 h. After 24 h, TNF-α, IL-6 and IL-12p70 production was analyzed by ELISA from culture supernatants. All data are expressed as mean ± SD (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001 for treatment compared to untreated controls (CON) or for difference between treatment data. n.s., no significant difference.