FIG 3.

FAO inhibition promotes NADPH oxidase recruitment on Mtb phagosomes. (A) Survival of Mtb in BMDMs from Nox2 KO and control mice that were untreated or treated with the indicated concentrations of ETM and TMZ, 120 hpi.*, P = 0.04; ****, P = 0.0001, ordinary one-way ANOVA. (B) Immunofluorescence (IF) microscopy of gp91^phox/NOX2 (green) and dsRed-expressing H37Rv (red) in BMDMs that were infected and treated with 5 nM TMZ or untreated for 24 h. The colocalized region is shown in yellow. Bars = 10 μm. (C and D) MFI of the NADPH oxidase subunits gp91^phox/NOX2 (C) and p40^phox (D) colocalized with H37Rv in BMDMs that were untreated or treated with 5 nM TMZ (24 hpi). (E) MFI of p40^phox colocalized with Mtb in BMDMs treated with 5 nM TMZ alone or in the presence of mitoTEMPO (48 hpi). (C to E) Automated image analysis was used to quantify gp91^phox/NOX2 or p40^phox MFI colocalized with more than 100 bacilli in at least two independent experiments. Data show means ± SEM of MFI from around individual bacilli in the sample. *, P = 0.02, ****, P ≤ 0.0001 calculated using unpaired Student’'s t test with Welch’s correction (C and D) or ordinary one-way ANOVA (E).