Flowchart of cryo-EM processing steps for the S. rosetta caveolin complex. (A) Flowchart depicting the classification and analysis of S. rosetta caveolin complex micrographs. Ab initio reconstruction used as an input for nonuniform refinement is shown in light blue in an en face view and rotated 90° around the x axis. Nonuniform and local refinements with no symmetry applied (C1) and 11-fold symmetry applied (C11) of the S. rosetta caveolin complex are shown in green in an en face view. (B) Representative micrograph of the S. rosetta caveolin complex. Scale bar, 50 nm. (C) Representative S. rosetta caveolin complex 2D classes. Box size, 300 pix² (261 × 261 Å). Scale bar, 100 Å. (D) GS-FSC of C11 refinement with no mask (blue line), loose mask (green line), tight mask (red line), and corrected mask (purple). Blue horizontal line, FSC = 0.143. (E) Euler angle plot of angles of particle distribution for the C11 reconstruction of the S. rosetta caveolin complex. (F) Heat map of local resolution of C11 3D reconstruction, rotated around the x axis. GS-FSC, gold-standard Fourier shell correlation; FSC, Fourier shell correlation.