Extended Data Fig. 7: Biochemical and structural characterizations of human MLL3–ubNCP complex.

a, Gel filtration (left) and SDS–PAGE analysis (right) of the assembly of the recombinant MLL3 catalytic module, composed of full-length WRAD proteins and MLL3 (residues 4707–4911). Experiments were repeated at least three times with similar results. b, EMSAs of the recombinant human MLL3 complex with either unmodified or H2BK120ub1 NCPs at molar ratios of 1:1, 2:1 and 4:1. Top, input of the EMSA mixtures. Bottom, native PAGE analysis of the gel shifting of NCP and ubNCP by the MLL3 complex. Each assay was repeated at least three times with similar results. c, Time course of the H3K4 methylation catalysed by the recombinant human MLL3 complex on unmodified and H2BK120ub1 NCPs. Error bars denote the s.d. from the mean of three replicates. d, Representative micrograph of the cryo-EM dataset of the MLL3–ubNCP complex. e, Representative 2D class averages of cryo-EM particles of the MLL3–ubNCP complex. f, Angular distribution of particle projections of the MLL3–ubNCP reconstruction. g, The gold-standard FSC curve calculated between two halves of the MLL3–ubNCP dataset. h, Local-resolution estimates of the structure of the MLL3–ubNCP complex.