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. 2021 May 4;12:666293. doi: 10.3389/fimmu.2021.666293

Figure 1.

Figure 1

Recombinant Rv2145c induces macrophage activation. (A) Rv2145c purified from E coli extracts was analyzed by SDS-PAGE with Coomassie blue staining and immunoblot staining using an anti-His antibody. (B) The cytotoxic effects of Rv2145c were analyzed by flow cytometry. BMDMs were stimulated with Rv2145c (1, 5 or 10 μg/ml), LPS (100 ng/ml) or staurosporine (STS, 100 nM) for 24 h and then stained with Annexin V and PI. The percentage of positive cells in each quadrant is indicated. (C, D) BMDMs were stimulated with Rv2145c (1, 5 or 10 μg/ml) or LPS (100 ng/ml) for 24 h. IL-10, IL-6, TNF-α, and IL-12p70 levels in the culture supernatants were measured by ELISA (C). The expression of surface markers was analyzed by two-color flow cytometry (D). 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 bar graphs show the percentage (mean ± SD of three experiments) for each surface molecule on F4/80+ cells. All data are representative of three experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 for treatment compared with medium controls (MC) or for the difference between treatment data. n.s., no significant difference.