Supplementary Figure 9: Data processing procedures for PRC2–DiNcl30
From: Cryo-EM structures of PRC2 simultaneously engaged with two functionally distinct nucleosomes
(a) Template-based particle picking from 1,532 micrographs resulted in selection of 190,479 particles. 3D classification and alignment yielded a major class of good particles (green box), which was further refined to obtain the final 3D reconstruction. (b) FSC curve of the PRC2-DiNcl30 refined dataset, indicating an overall resolution of 8.4 Å according to the FSC = 0.143 criterion (Rosenthal and Henderson, J. Mol. Biol. 333, 721-745, (2003), Scheres and Chen, Nat. Methods 9, 853-854, (2012)). (c) Angular distribution of particle orientations after refinement. (d). Map of the PRC2-DiNcl30 complex colored according to local resolution ranging from 9-13 Å. Local resolution estimation determined with BLOCRES (BSOFT package (Heymann and Belnap, J. Struct. Biol. 157, 3-18, (2007)). (e) Detailed views of the SANT1/SBD and the EED/SBD contacts with the nucleosomal DNA in the PRC2-DiNcl30 complex viewed from the front (left) and back (right) of the complex. Red dot, possible SANT1-EED contact as in Fig. 4d. Asterisks, EED-DNA contacts, coloring according to Fig. 4. Green dot, linker segment connecting SANT1 and the SRM.
