IncuCyte Zoom Live-Cell Imaging.

PMA-differentiated THP-1 cells were seeded on 24-well dishes at either 400,000 or 100,000 cells per well. Cells were treated with 100 nM Sytox (Sytox) (S7020; Thermo Fischer) to assay cell death and/or permeabilization or in separate wells with Syto-Green (S7559; Thermo Fischer) to assay total cell number. Cells were imaged with the IncuCyte imaging platform (Essen Bioscience). At each time point, four images were taken per well. Each treatment was run in either duplicate or triplicate. Percent cell death was calculated by averaging Syto-Green counts over 2 h near the beginning of the run. These were averaged with replicates and used to normalize Sytox counts to calculate percent cell death. For quantitation of cell swelling, IncuCyte images were imported into ImageJ/FIJI, and area measurements were made on >50 cells from a single field of view for each condition.