Fig. 2. Cryo-EM workflow for inhibitor series structure determination.

a Chemical structures of pyrazolopyrimidine-type inhibitors used for cryo-EM structure determination. The chemical core shared by all inhibitors is shown in the first position, along with definition of the R₁-R₃ substituent positions that will be used in the text. b Schematic of tripartite cryo-EM workflow. The possibility of using a two-stage workflow is indicated by a dashed green arrow. c Example result from 1-h Glacios screening with visible ligand density (purple). d Example result from 4-h Glacios screening with visible ligand density (purple). e Example high-resolution cryo-EM map from Krios G4 collection with visible ligand density (purple).