Extended Data Fig. 2 |. Cryo-EM data processing for the WT GSDMD dataset.

a, A cryo-EM image of the WT GSDMD sample. Scale bar: 100 nm. b, Processing of the WT GSDMD cryo-EM dataset. Initial 2D classes showed a ring-stacking phenomenon, which added to structural heterogeneity and posed challenges to symmetry determination. Density subtraction was therefore performed, followed by 3D reconstruction of each ring without assumption of symmetry, after which particle symmetry could be determined for certain 3D classes. These classes were then refined with their respective symmetry imposed to yield final cryo-EM maps. Data shown in a are representative of three independent experiments.