Figure 7

ESAT-6 mediates mitochondrial damage in mTOR-deficient macrophages downstream of its involvement in phagosomal permeabilization

(A–E) Torin1- and DMSO-treated THP-1 macrophages were infected with tdTomato-expressing WT or ΔESX-1 Mm at MOI = 3 and treated with prazosin (PRZ, 20 μM) for 7 h. See also Figure S4.

(A) Confocal micrographs of galectin-8 (GAL8) immunofluorescence (green) and Mm fluorescence (magenta) in THP-1 macrophages 7 hpi. GAL8 foci associated with Mm (filled arrowheads) or not associated with Mm (open arrowheads) are shown. Scale bar, 20 μm.

(B) Percentage of macrophages with GAL8-associated Mm foci.

(C) Percentage of Mm volume associated with GAL8 foci 7 hpi.

(D) Percentage of cells that have released cytochrome c 7 hpi.

(E) mtor^{sa16755/sa16755} fish and mtor-sufficient siblings were infected with ∼90 fluorescent Mm via the hindbrain ventricle on 2 dpf. On 1 and 2 dpi, animals were injected with ∼3 nL of 300 μM PRZ or 1% DMSO into the hbv. Graph indicates the percentage of animals with cording 3 dpi.

(F) Wild-type fish treated with 400 nM rapamycin were infected with ∼180 tdTomato-expressing ΔesxA Mm complemented with WT or point mutant Mtb esxA via the hbv on 2 dpf. Animals were injected with PRZ or DMSO as indicated on (E). Graph indicates the percentage of animals with cording 3 dpi.

Symbols represent values from individual (B and C) imaging fields or (D) individual wells. (B and C) Horizontal lines and (D) bars indicate mean values. (E and F) Numbers within columns indicate animals per group. Statistical analyses, (B and D) one-way ANOVA with Sidak’s post-test or (E and F) Fisher’s exact test.

See also Figure S4.