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 5. miR-1281 inhibition upregulates CypD and intensifies MTB-induced cytotoxicity in human macrophages. The primary human macrophages were transduced with the lentiviral pre-miR-1281 anti-sense (“antagomiR-1281”) or the non-sense control miR anti-sense (“antagomiR-C”), with stable cells selected by puromycin. Macrophages were then infected with Mycobacterium tuberculosis (MTB) for applied time periods, expression of mature miR-1281 and listed genes was shown (A and C); The relative CypD 3’-UTR activity was tested (B); Mitochondrial depolarization, cell viability, cell necrosis and apoptosis were tested by JC-1 staining (D), CCK-8 (E), medium LDH release (F), and caspase-3 activity (G)/TUNEL staining (H) assays, respectively. CypD expression was quantified, normalized to Tubulin (C). Data were presented as mean ± SD (n=5). * P <0.05 vs. “C” treatment in “antagomiR-C” macrophages. #P <0.05 vs. MTB treatment in “antagomiR-C” macrophages. Experiments in this figure were repeated five times with similar results obtained.