Lactate dehydrogenase (LDH) assay.

LDH released from pyroptotic cells was measured by a Cytotoxicity Detection LDH Kit (Roche, 11644793001). In brief, 10,000 BMDMs per well were seeded in a 96-well plate and treated with or without LPS transfection. For LDH release detection, 100 μL supernatant was collected from each well. After centrifugation (4°C, 350g, 5 minutes), 100 μL of the kit’s reaction mixture were added and coincubated for 30 minutes in the dark. The microplate reader (Molecular Devices, Spectra Max Plus) was used to measure the OD at 490 nm. The final LDH release was calculated according to the manufacturer’s protocol.