Griess assay

BMMs were plated in 96‐well plates at 100 000 cells per 100 µL overnight, after which cells were pretreated with mitochondrial fission inhibitors M1 (20 µm) and mdivi1 (10 µm) for 1 h, prior to stimulation with LPS (100 ng mL⁻¹) for the indicated time points. To induce IL‐1β secretion, BMMs were stimulated for 4 h with LPS, followed by treatment with nigericin (10 µm; Sigma, St. Louis, MO, USA) for 4 h. To assess nitric oxide production, BMMs were pretreated with IFNγ (5 ng mL⁻¹; R&D System, Minneapolis, MN, USA) for 18 h, after which cells were stimulated with LPS for 24 h. Nitrite levels were measured by Griess assay (Promega, Madison, WI, USA) and levels of secreted Il‐12p40, Il‐6, Il‐1β and Tnf were measured by ELISA according to the manufacturer’s instructions [TNF and IL‐1β kits: BD Biosciences, Franklin Lakes, NJ, USA; IL‐6 antibodies: BD Biosciences (Capture: #554400, Detection: #554402); IL‐12p40 antibodies (Capture: #551219, Detection: #554476)].

Levels of human IL-6, IL-10, IL-12, and TNF-α were quantified using ELISA kits from BioLegend, Levels of mouse IL-6, TNF-α, and IL-10 were determined using ELISA kits from BioSite (Täby, Sweden). Nitrite levels were measured using a Griess assay (Promega). All analyses were performed according to the manufacturer’s instructions.