FIGURE 7.

Induction of autophagy in peripheral blood mononuclear cells infected with Mycobacterium tuberculosis H37Rv-mCherry and treated with simvastatin. A total of 2 × 10⁶ PBMCs was seeded in a 24-well plate and cultured under the following conditions: (1) PBMCs infected at an MOI of 1 with M. tuberculosis strain H37Rv-mCherry, and (2) cells infected at an MOI of 1 with M. tuberculosis strain H37Rv-mCherry and treated with 20 μM simvastatin acid. Uninfected no treated cells were included as controls. The cells were incubated at 37°C in a 5% CO₂ atmosphere for 24 h and then stained with Cyto-ID and Hoechst 33342 for detection of autophagosomes (green) and visualization of nuclei (blue), respectively. The stained cells were analyzed using confocal microscopy. (A) Intact mycobacteria were observed inside some infected PBMCs (red). The induction of autophagy was also observed independent of infection (green). (B) Sequestration of red stain within autophagosomes (green) and loss of the integrity of mycobacteria were observed in infected and treated PBMCs. Scale bar, 5 μm.