Figure 4.

mTOR deficiency promotes mycobacterium-induced, mitochondrially mediated cell death

(A and B) THP-1 macrophages were infected with (A and B) tdTomato- or (C and D) BFP-expressing Mm at MOI = 3.

(A) Flow cytometry histograms of cytochrome c (cyt c) fluorescence in infected viable cells (FVD eFluor 660⁻) 7 h post-infection (hpi). Gate indicates cells that have released cyt c.

(B) Quantification of cyt c^low cells 7 hpi.

(C and D) Torin1-treated THP-1 macrophages were labeled with TMRE and MitoTracker Deep Red prior to imaging in the presence of Sytox Green 32 hpi. See Video S3 and Figure S2.

(C) Confocal micrographs of a dying infected macrophage (filled arrowhead) surrounded by surviving uninfected macrophages. Mm (asterisk), Sytox Green (open arrowheads). Scale bars, 10 μm.

(D) MFI of TMRE, MitoTracker Deep Red, and Sytox Green staining of dying infected macrophages over time. Key time-lapse frames for cell 1 are shown in (C).

Statistical analyses, (B) two-way ANOVA with Tukey’s post-test.