Measurement of H2O2 production
The supernatants of stimulated peritoneal murine macrophages cultures were collected and analyzed for hydrogen peroxide (H2O2) concentrations by using a colorimetric assay adapted to the laboratory conditions36,39. Briefly, 50 μL supernatants from each culture were placed into a 96-well plate and mixed an equal volume of fresh phenol red solution (5.5 mM dextrose, 0.056 g phenol red and 8.5 U/mL Type I HRP in DPBS) followed by incubation at room temperature and protected from light for 3 h. Afterwards, the reaction was stopped by adding NaOH solution, and absorbance was measured at 490 nm using a microplate absorbance reader (Thermo Scientific, Multiskan FC). Hydrogen peroxide concentration was determined based on a H2O2 standard curve ranging from 0 to 50 μM/mL. All experimental variants were performed in triplicate.