Reanalysis of cryo-EM data reveals ALK-cytokine assemblies with both 2:1 and 2:2 stoichiometries
Fig 3
Comparison of map (an)isotropy following data processing strategies with or without Orientation Rebalancing and/or 3D refinement with enabled Blush Regularization.
(A) Processing workflows (1–5) of cryo-EM dataset EMPIAR-10930 starting from 876,806 selected particles as outlined in Methods. Workflow 1 does not include Orientation Rebalancing (CryoSPARC v4.5) nor 3D refinement with Blush regularization (RELION v5). Workflow 2 is the same as 1 but with a final 3D refinement with Blush regularization in RELION v5 instead of NU-refinement in CryoSPARC v4.5. Workflow 3 utilizes Orientation Rebalancing in CryoSPARC v4.5 followed by NU-refinement in CryoSPARC. Workflow 4 includes Orientation Rebalancing in CryoSPARC v4.5 followed by regular 3D refinement in RELION (without Blush regularization), while workflow 5 combines Orientation Rebalancing in CryoSPARC with 3D refinement with enabled Blush regularization in RELION. (B) Conical FSC summary plots for the final 3D refinements obtained in processing workflows 1, 2, 3, 4, and 5, generated via “Orientation Diagnostics” in cryoSPARC v4.5. The resolution at FSC = 0.143 is annotated in blue.