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 1. MTB infection induces mPTP opening and programmed necrosis in human macrophages. The primary human macrophages were infected with Mycobacterium tuberculosis (MTB) for applied time periods, mitochondrial immunoprecipitation (Mito-IP) assays were carried out to test CypD-ANT1-p53 association in the mitochondria (A, “Mito-IP”), with expression of these proteins examined by Western blotting (A, “Input”); Mitochondrial depolarization was examined by JC-1 dye assay (B); Cell necrosis was tested by medium LDH release assays (C). For JC-1 assays, both JC-1 merged images and JC-1 green fluorescence intensity were presented (same for all Figures). Expression of listed proteins was quantified, normalized to loading controls (A). “C” stands for uninfected control macrophages (same for all Figures). Data were presented as mean ± SD (n=5), and results were normalized to “C”. * P <0.05 vs. “C” macrophages. Experiments in this figure were repeated five times with similar results obtained. Bar= 100 μm (B).