Extended Data Fig. 3 |. Cryo-EM data processing for the L192E GSDMD dataset.

a, A cryo-EM image of the L192E GSDMD sample. Scale bar: 100 nm. b, Processing of the L192E GSDMD cryo-EM dataset. The L192E dataset was first processed following the procedures for the WT dataset. Cryo-EM maps obtained from 3D refinement with symmetry imposed were further classified without alignment to remove heterogeneous particles. Then, the best 3D classes were refined again to improve resolutions. A 3D reconstruction at 7.3 Å was further improved by symmetry expansion, 3D classification without alignment, 3D local refinement, and per-particle CTF refinement to reach the final map at 3.9 Å resolution. c, Similarity of cryo-EM maps generated from the WT and L192E datasets. Due to the higher resolutions, maps from the L192E dataset were chosen for model building. Data shown in a are representative of three independent experiments.