Fig. 4

Mitochondria undergo oxidative stress during infection. (A) mtROS production in infected hMdM was measured via flow cytometry. The results of four biological replicates are shown. Significance was calculated with the Mann-Whitney U test (* p ≤ 0.05). (B) Mitochondrial ATP was quantified using flow cytometry. Significance was calculated with the Mann-Whitney U test (* p ≤ 0.05). (C) Mitochondrial mass in hMdM after infection was measured via staining with MitoTracker Green. The results of six biological replicates are shown. Significance was calculated with the Mann-Whitney U test (ns p ≥ 0.05). (D) Mitochondrial membrane potential was quantified by staining with MitoTracker Red and measurement via flow cytometry. Significance was calculated with the Mann-Whitney U test (ns p ≥ 0.05). (E) Heatmaps showing individually differentiated genes related to regulation of mitochondrial metabolism, (F) imported mitochondrial proteins and (G) regulation of mitochondrial homeostasis. Gene expression is presented over log2 fold change. At least one DEG per gene per condition had a significant log2 fold change. (H) Schematic representation of THP-1 redox cell model showing subcellular location of different roGFP2 constructs. (I) Determination of redox status of THP-1 cells stably expressing cytosolic roGFP2, roGFP2 coupled to mitochondrial ATPase and OTC. Redox status was analyzed by measuring fluorescence at λex 405 nm and 488 nm. RFI is normalized to control cells. Significance was calculated with Kruskal-Wallis test (* p ≤ 0.05). (J) Immunofluorescence staining of infected hMdM stained with MitoTracker Red, phalloidin and DAPI