Fig. 3

TBCM regulates apoptotic cell death in macrophages. BMDMs were isolated from female BALB/c mice and infected with BCG or BΔtbcm at an MOI of 10. A Apoptotic cells were detected by the TUNEL assay at 24 h postinfection. B Early or late apoptotic cell death was quantified by the Annexin V/PI assay at 24 and 48 h postinfection, and the statistical significance is shown in the table. C The expression level of cleaved caspase-3 was analyzed by western blotting at 18 and 24 h postinfection. D The mRNA expression levels of apoptosis-associated genes were measured by real-time PCR. The Bcl-2, caspase-7, caspase-9 and Apaf-1 mRNA expression levels were measured in infected BMDMs at 2 and 4 h or 12 and 24 h postinfection. All data were collected in at least three independent experiments and are presented as the mean ± SEM. The results were analyzed for statistical significance by two-way ANOVA with Tukey’s multiple comparison test. (*P < 0.05, **P < 0.01, ***P < 0.001). BCG, wild-type BCG; BΔtbcm, BCG Δtbcm mutant