Fig. 6.

Complementation of the tbcm gene restored the inhibition of intrinsic apoptosis. J774A.1 cells were infected with BCG, BΔtbcm or CΔtbcm at an MOI of 10. A Apoptotic cells were quantified by the Annexin V/PI assay at 48 h postinfection, and the statistical significance is shown in the table. B Bcl-2, caspase-7, caspase-9 and Apaf-1 mRNA expression levels were measured by real-time PCR at 0, 4 h or 12 and 24 h postinfection. C Expression levels of BAX and BAK were detected in the mitochondrial fraction by western blotting. D Cytochrome c in the cytosolic fraction was detected at 6 h postinfection by western blotting. E Cytosolic cytochrome c was measured by ELISA at 8 h postinfection. F Cells were stained with JC-1 dye at 24 h postinfection. G Restored TRIM39, SARM1, SLC25A26, NUFA8 and DTYMK gene expression levels were confirmed after tbcm complementation at 0, 2 and 4 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 (A, B, G) or one-way (C, D, E) ANOVA with Tukey’s multiple comparison test. (*P < 0.05, **P < 0.01, ***P < 0.001). BCG, wild-type BCG; BΔtbcm, BCG Δtbcm mutant; CΔtbcm, complemented mutant