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. 2015 Nov 6;5:16320. doi: 10.1038/srep16320

Figure 1. Virulent Mycobacterium tuberculosis selectively inhibits autophagy flux.

Figure 1

PMA differentiated (20 ng/ml, 24 hours) THP-1 macrophages were infected with PKH67 (green) labelled H37Ra or H37Rv at MOI of 1:10 (methods). Autophagosomes were visualized by LC3 immuno-staining at 6, 12, 24, 48 and 72 hours post infection with and without BafA1 treatment (100 nM, 3 hr). (A) Representative images showing co-localization of both H37Ra and H37Rv (green) to autophagosomes (LC3, red) at 48 hours post infection. Panels (B,C) show Co-localization Coefficient M1 of Mtb and LC3 for H37Ra and H37Rv infection, respectively with and without BafA1 treatment (Mean ± Standard Error Measurements, *p < 0.05). Panel (D) shows representative images of Mtb-infected mouse BMDMs with LC3 (red) at 48 hours post infection with and without BafA1. Panel E shows the corresponding quantification of the dataset (mean ± SEM, **p < 0.01). Panel (F) shows LC3 immunoblots from the cell lysates of H37Ra or H37Rv infected macrophages as well as uninfected control macrophages at 6 and 48 hours post infection. Cells were treated with BafA1 (100 nM, 3 hr) to assess autophagy flux. Panel (G) shows representative images of Mtb in autolysosomes (LC3:LysoTracker; (blue:red)) at 48 hours post infection. The plot for the same at 6, 12, 24, 48 and 72 hours post infection is depicted in Panel (F) (Mean ± Standard Error Measurements, *p < 0.05).