Fig. 6.
Cell swelling precedes pyroptotic cell rupture. a–d B6Nlrp1b+ or B6 BMDMs stained with Cholera Toxin subunit B-Alexa 594 (rCTB) were stimulated with LeTx (a, b) or FlaTox (c, d), respectively, and imaged in culture media containing Sytox Green. Confocal images were acquired every 1.5 min. Graphs show the percentage of mean fluorescence intensity (MFI) of Sytox Green and cell volume quantifications based on rCTB-Alexa 594 staining, both calculated as described in the Methods section. Values represent the mean ± SD of individual cells imaged in 3 independent experiments (LeTx, n = 26; FlaTox n = 16). Fluorescent micrographs show the maximum intensity projection (Sytox Green) or the single plane (rCTB) of a representative cell. Single-cell plots are shown in Supplemental Fig. 7. In all panels, time point zero indicates the first detection of Sytox Green. All scale bars, 10 µm