Extended Data Fig. 9: Supporting data for cryo-EM investigation of H. sapiens replisome: CUL2^LRR1 complexes.

a, Schematic of reconstitution approach used for preparation of replisomes bound to CUL2^LRR1 for cryo-EM. A schematic of the DNA substrate used is shown with a 39 nucleotide 3′ arm and no 5′ arm. b, Silver-stained SDS-PAGE gels analysing 100 μL fractions taken across 10-30% glycerol gradients, either lacking (top) or containing (bottom) crosslinking agents. Fractions 15+16 used for cryo-EM sample preparation are indicated. This experiment was performed twice. c, Representative cryo-EM micrograph. d, Representative 2D class averages, 40 nm box width. e–j, (Top) cryo-EM reconstructions coloured by local resolution according to inset keys (Bottom) angular distribution of particle orientations. e, Consensus refinement for replisome:CUL2^LRR1 fully engaged. f, Consensus refinement for replisome:CUL2^LRR1 where the LRR1^PH domain is bound but the LRRs are disengaged. g, Consensus refinement for particles lacking CUL2^LRR1. h, Multi-body refinement for AND-1:CDC45:GINS. i, Multi-body refinement for LRR1:ELOB:ELOC:CUL2:AND-1-HMG. j, Multi-body refinement for CUL2:RBX1. k, Cryo-EM density for the LRR1 LRRs. l, Cryo-EM density for the LRR1 PH domain. m, Representative cryo-EM density for a LRR1 LRR domain β-strand at 3.5 Å resolution. n, Representative cryo-EM density for a LRR1 LRR domain α-helix at 3.7 Å resolution. o, Fourier-shell correlation (FSC) curves for the various maps used in model building. p, Map-to-model FSC curves for the complete model docked into the consensus refinement for replisomes fully engaged by CUL2^LRR1. For gel source data, see Supplementary Fig. 1.