Dose-fractionated movie stacks were gain-corrected, aligned, and dose-weighted using IMOD (44), and motion-corrected using MotionCor2 (45) within RELION 3.0 (46). Gctf (47) version 1.06 was used for estimation of defocus. About two thousands particles were selected by manual picking, and 2D classes were constructed from these in RELION 3.0; these classes were then used as templates for autopicking of all micrographs, yielding 213,528 picked particles. Two rounds of 2D classification of these particles were performed, and well-resolved classes were chosen yielding 178,666 particles. An initial ab initio model was created using RELION 3.0’s “InitialModel” function. The “Refine3D” function was used to create a higher-resolution model of the density map using the initial model as a template. Masking and postprocessing in RELION 3.0 resulted in a global resolution of 3.5 Å without applying symmetry. The contrast transfer function values were refined, and Bayesian polishing was performed in RELION 3.0. The final particle dataset was refined using C3 symmetry. After masking and postprocessing, the global resolution was determined using the gold-standard Fourier shell correlation of 0.143 cutoff criterion to be 3.19 Å (fig. S2) (46, 48).

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