Figure 2.

Mtb infection of IFN-γ-stimulated macrophages with the Δami1 and Δami4 mutant induced elevated proinflammatory responses with no effect on macrophage cell viability and intracellular Mtb growth. Cell viability of IFN-γ-stimulated (100 U/mL for ~12 h) BMDM infected with the (a) Δami1 mutant, complemented strain (Δami1::ami1) and wild-type H37Rv and (b) CFU counts of BMDM infected with the Δami1 mutant, complemented strain (Δami1::ami1) and wild-type H37Rv. Proinflammatory cytokine production from infected macrophages was measured via ELISA at 4- and 6-days post-infection for BMDMs infected with the (c) Δami1 mutant, complemented strain (Δami1::ami1) and wild-type H37Rv. (d) Cell viability of IFN-γ-stimulated (100 U/mL for ~12 h) BMDM infected with the Δami4 mutant, complemented strain (Δami4::ami4) and wild-type H37Rv was measured via Cell Titer Blue at 0 (4 hours), 2, 4 and 6 days post-infection. (e) CFU counts of BMDM infected with the Δami4 mutant, complemented strain (Δami4::ami4) and wild-type H37Rv. (f) Proinflammatory cytokine production from infected macrophages was measured via ELISA at 4- and 6-days post-infection for BMDMs infected with the Δami4 mutant, complemented strain (Δami4::ami4) and wild-type H37Rv. (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, one-way ANOVA, n = 3). DPI = days post-infection. Data are representative of one (1A, 1D) and two (1B, 1 C, 1E and 1 F) experiments