Research Paper Volume 11, Issue 24 pp 12661—12673

Targeting cyclophilin-D by miR-1281 protects human macrophages from Mycobacterium tuberculosis-induced programmed necrosis and apoptosis

Figure 4. miR-1281 overexpression inhibits MTB-induced programmed necrosis and apoptosis in human macrophages. The primary human macrophages were infected with lentivirus encoding pre-miR-1281 (“lv-pre-miR-1281”), two stable cell lines, “Line1/2”, were established following puromycin selection. Control macrophages were infected with non-sense microRNA (“lv-C”); The macrophages were infected with Mycobacterium tuberculosis (MTB) for applied time periods, mitochondrial depolarization, cell viability, cell necrosis and apoptosis were tested by JC-1 staining (A), CCK-8 (B), medium LDH release (C), and caspase-3 activity (D)/TUNEL staining (E) assays, respectively. Data were presented as mean ± SD (n=5). * P <0.05 vs. “C” treatment in “lv-C” macrophages. #P <0.05 vs. MTB treatment in “lv-C” macrophages. Experiments in this figure were repeated five times with similar results obtained. Bar= 100 μm (A and E).