FIG 1.

BMDMs from huTNF KI mice respond to BCG, LPS, and IFN-γ as well as WT cells producing cytokines, chemokines, and phosphorylated NF-κB p65 upon BCG infection. BMDMs from huTNF KI and WT mice were activated for 24 h with BCG Connaught (MOI, 1), LPS, or IFN-γ. (A) Murine TNF concentrations in the supernatants of WT BMDMs and human TNF concentrations in the supernatants of huTNF KI BMDMs. (B to E) The levels of soluble TNFR1 (B), soluble TNFR2 (C), IL-6 (D), and MCP-1 (E) in culture supernatants were determined (n = 6). NT, not treated. (F) Phosphorylated and nonphosphorylated NF-κB p65 proteins in BMDMs uninfected or infected with BCG were detected by Western blotting at different time points. One of three representative Western blots is shown. (G) The graph presents the ratio of phosphorylated NF-κB p65 (P-NF-κB) to nonphosphorylated NF-κB p65 bands from three Western blots corresponding to three experiments. Quantification was done with Quantity One analysis software, and results are presented as the ratio of the number of units of phosphorylated NF-κB p65/number of units of total NF-κB p65 (n = 3 mice per group). The bar graphs show means ± SEMs. *, P < 0.05 versus the WT.