Extended Data Fig. 5: Single-particle cryo-EM image processing workflow for the SprA complexes.

Cryo-EM data sets for SprA complexes in LMNG, in the presence or absence of fluorinated detergents, were combined following 2D classification and subjected to 3D classification against a low-resolution model generated from the fluorinated octyl-maltoside data set. Particle images corresponding to the PorV complex or the Plug complex were then independently refined. A soft mask, generated from these maps, was then used to perform masked refinement against the same particle images, resulting in global map resolutions of 3.5 Å for the PorV complex and 3.7 Å for the Plug complex.