Extended Data Figure 5. Flow chart of cryo-EM data processing and resolution analysis of Rho*–GRK1S5E/EE−Fab6.
a) A representative raw cryo-EM micrograph from a total of 8,358. From these, 7.5 million particles were automatically picked in cryoSPARC and were used to generate 2D class averages (shown are representative good classes). After screening out bad classes, the remaining 353 thousand particles were further processed using heterogeneous refinement. Class 2 and Class 3 showed similar quality and were selected for homogeneous refinement and then non-uniform refinement in cryoSPARC. The resolution of the final map is estimated to be 4.0 Å. Global resolution was determined by FSC with a cut-off of 0.143. b) Directional FSC indicates that the resolutions in the x and z directions are similar to the globular resolution, whereas the resolution in the y direction was lower. c) Plots of the global FSC together with the spread of directional resolution values defined by ±1 σ from the mean and a histogram of 100 such values evenly sampled over the 3D FSC. d) Local resolution map as estimated by cryoSPARC.