Fig. 5: Deletion of MMAR_0267 inhibited the level of apoptosis.

a THP-1 cell RNA was extracted to measure the expression levels of cytokines P53, BCL2, Cas9, and BAX. Error bars represent standard deviation; (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001; n = 3). b Immunoblotting was used to detect apoptosis-related molecules in THP-1 macrophages. Protein quantification of P53, BCL2, and Cas9 was conducted using ImageJ software. Western blot results were based on three replicates with triplicate samples for each condition. Grayscale analysis was utilized for quantification and statistical analysis (n = 3). c THP-1 macrophages were infected with WT-MM, ΔMMAR_0267, and ΔMMAR_0267::MMAR_0267 strains labeled with GFP at an MOI of 20:1. Co-localization was analyzed using confocal immunofluorescence microscopy with LysoTracker-Blue DND staining. Representative images show saphirine representing co-localization of fluorescent signals. Scale bars are 20 μm. d Quantification of bacterial co-localization with LysoTracker DND blue. Mean values ± 1 s.e. depict the percentage of bacteria co-localizing with the LysoTracker signal in ~100 random cells, analyzed in a blinded fashion. Statistical significance: *P < 0.05, **P < 0.01 for WT-MM, ΔMMAR_0267, and ΔMMAR_0267::MMAR_0267 (two-tailed Student T test).