Extended Data Fig. 8: CryoSPARC workflow for structural determination of the FlaA, FlaB, and FlaC filament in V. cholerae.
From: Structures of the sheathed flagellum reveal mechanisms of assembly and rotation in Vibrio cholerae
(a) 15,919 micrographs of ΔflhG V. cholerae were collected to determine the FlaA filament structure. Scale bar: 100 nm. (b) Representative 2D classifications of the hook–filament junction region after manual picking and Topaz picker30. Scale bar: 200 Å. (c, d) Cryo-EM density map of the hook–filament junction with FlaA filament, resolved at 3.43 Å based on the “gold standard” FSC0.143 plot. (e) Snapshot of manual picking at the filament tip region. Scale bar: 100 nm. (f) Representative 2D classifications of filament tip particles from 19,040 manually picked particles. Scale bar: 200 Å. (g, h) Cryo-EM density map of the FlaB filament at the filament tip, resolved at 3.16 Å. (i) Snapshot of filament tracer in ΔflhGΔflaBDE V. cholerae. Scale bar: 100 nm. (j) Representative 2D classifications of the FlaC filament. Scale bar: 200 Å. (k, l) Cryo-EM density map of the FlaC filament in ΔflhGΔflaBDE V. cholerae, resolved at 3.16 Å.
