Figure 3.

NO^● and O₂^●− donors inhibit the respiration of naïve macrophages. Macrophages were activated for 24 h with the IFNγ + LPS (40 ng/mL and 10 ng/mL respectively) or not (untreated) and subjected to serial injections of oligomycin (2 μΜ), FCCP (1 μΜ) and antimycin A/rotentone (2/2 μΜ), according to the mitostress protocol. A Seahorse XFe96 metabolic analyzer was used to record the changes in the oxygen consumption rate (OCR). The reactive species-donors SIN-1 (NO^● and O₂^●−) or SNAP (NO^●) were added (3 mM for either) to untreated (naïve) macrophages prior to the assay. For comparison reasons horizontal intermittent line indicate the basal OCR levels of IFNγ + LPS activated macrophages. The vertical intermittent lines indicate the injection points. One representative experiment is shown, with data presented as mean ±STD of at least three replicates per condition.