Fig. 7

The autophagy-related gene ATG5 controls mitophagy. A RAW 264.7 cells were infected with Mtb (MOI = 1) for 24 and 48 h. B–H WT and BNIP3-KO RAW 264.7 cells were transfected with siRNA and then infected with Mtb (MOI = 1) for 48 h. B Mitophagy was measured in RAW 264.7 cells using a mitophagy detection kit (n = 4 per group). C MMP was measured in RAW 264.7 cells using JC-1 staining. Fluorescent intensity of JC-1 was detected at excitation wavelength 488 nm and emission wavelength 530 nm using flow cytometry (n = 4 per group). D WT and BNIP3-KO RAW 264.7 cells were incubated with MitoSOX Red to measure mROS levels. The mROS levels were analyzed using flow cytometry (n = 4 per group). E The analysis of proinflammatory cytokine levels was performed in macrophages at 48 h post infection (n = 6 per group). F Intracellular survival was assayed by enumerating the CFU (n = 9 per group). G WT and BNIP3-KO RAW 264.7 cells were transfected with siRNA and then infected with Mtb (MOI = 1) for 48 h, stained with LC3 (green), Tom20 (red), and DAPI (blue), and visualized using confocal microscopy. H Quantification data in the Fig. 7G. Quantification data of LC3 recruitment to mitochondria were collected from 50 to 60 cells from at four independent experiments. The data are presented as means ± SD of at least three independent experiments; *p < 0.05, **p < 0.01 ***p < 0.001