BMDMs were seeded on 96-well (RodTox stimulation) or 24-well tissue culture plates (nigericin stimulation). Before live-cell imaging, medium was replaced with fresh macrophage medium containing 1:5000 SYTOX Green dye (Life Technologies) and Hoechst 33342 nucleic acid stain (Molecular Probes). Cells were incubated at 37°C for 20 min before adding 5 μM nigericin or RodTox (20 ng/ml). After the stimuli treatment, cells were immediately visualized for SYTOX Green uptake (cell death) and Hoechst nuclei staining (total cell number) using Cytation 3 Imaging reader (BioTek). Images were taken every 2 min (nigericin) or 6 min (RodTox) for up to 30 min (nigericin) or 54 min (RodTox). Cell death percentage was calculated as the number of SYTOX Green–positive cells divided by the Hoechst-positive cell number.

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