Fig. 4: Asymmetric rupture preserves filopodia and corpse remnants.

a, Lattice light-sheet imaging of macrophage expressing PLCδ-PH-GFP (green) and incubated with 1 µM SiR-actin (magenta). Primed macrophages were stimulated with nigericin for the time indicated in each panel. Cells in examples (i) and (ii) are displayed on a shifted axis (indicated in top right of example (i)) to highlight the 3D morphology of macrophages undergoing pyroptosis. Images were processed in Arivis 4D and represented as volume renderings. Each channel was nonlinearly adjusted (gamma, 2) to represent cell features across a wide dynamic range. b, (i) LLAMA segmentation⁴⁶ of the SiR-actin signal of lattice light-sheet imaging, analyzing nigericin-stimulated cells. Cell body (green), filopodia (magenta) and pyroptotic blisters (blue) were segmented by machine learning and are displayed as volumetric projections. b, (ii) Line scans (indicated in yellow on the rotated MIP panel) relative to (i) are displayed as orthogonal slice views to represent the spatial distribution of pyroptotic cell features. Orthogonal slices were processed using Fiji. c,d, SEM of macrophage corpses produced by NLRP3- or CASP11-activating stimuli (nigericin, c; cytosolic LPS, d). Scale as indicated and magnified insets are to the right of each panel. e, Quantification of the cell and corpse area of WT LPS-primed BMDM stimulated with nigericin with or without EGTA. Phalloidin-labeled footprints of intact cells containing ASC specks (left), and cells that had undergone PMR and lost the bulk of cell mass (right) were hand-segmented in Fiji. Examples are included in Extended Data Fig. 5c. The area of thresholded phalloidin staining was quantified per cell, and analyzed by an unpaired two-sided Mann–Whitney test. f, Schematic view of F-actin remodeling to generate filopodia-rich corpses via GSDMD-dependent Ca²⁺ influx. Scale bars, 10 µm except where indicated otherwise, and data are representative of three independent biological replicates. Violin plots show mean (solid line) and first and third quartiles (dotted lines). Statistical significance ****P ≤ 0.0001.