Cell Culture Protocols for Various Cell Lines

All cell lines were maintained at 37 °C, 5% CO₂. HeLa cells (ATCC (Manassas, VA, USA) CCL-2) were cultured in DMEM (Corning, Corning, NY, USA) supplemented with 10% Equafetal bovine serum (Atlas Biologicals, Fort Collins, CO, USA) and 1× l-glutamine (D10). 3T3 cells (ATCC, CRL-1658) were cultured in D10 supplemented with 1 mM sodium pyruvate (Corning) and 1× non-essential amino acids (GE Healthcare, Pittsburgh, PA, USA). NK92MI cells (ATCC, CRL-2408) were cultured in Alpha MEM (GE Healthcare) supplemented with 10% Equafetal bovine serum, 1× l-glutamine, 0.2 mM myo-inositol, and 0.02 mM folic acid. Caspase 1/11^−/− macrophages were isolated from C57BL/6 mouse bone marrow and cultured as previously described [4 (link)]. BMDM were differentiated for 7–21 days prior to experiments in DMEM that was supplemented with 30% L929 cell supernatant, 20% premium fetal calf serum (VWR Seradigm, Radnor, PA, USA), 1 mM sodium pyruvate, and 1× l-glutamine.