Extended Data Fig. 3: The role of H3K4me2 in HR factors recruitment at centromeric DSBs in G1 and G2.

a, Quantification of SETD1A mRNA by RT-qPCR in NIH3T3 cells depleted of SETD1A (siSETD1A), normalized to GAPDH and expressed as relative to control (siSCR). b, Western blot analysis of LSD1 and tubulin in NIH3T3 cells expressing GFP-dCas9-LSD1 and control cells. c, Chromatin immunoprecipitation analysis of H3K4me2 enrichment at centromeres over the Input and relative to the no-antibody control in siSETD1A vs siSCR NIH3T3 cells. d, Chromatin immunoprecipitation analysis of H3K4me2 enrichment at centromeres over the Input and relative to the no-antibody control in cells expressing dCas9 or dCas9-LSD1 and a gRNA targeting the minor satellite repeats (mi gRNA). e, Immunofluorescence confocal analysis of NIH3T3 cells expressing Cas9 + mi gRNA and dCas9 or dCas9-LSD1 and stained with DAPI and antibodies specific for γ-H2AX, 53BP1, RPA, BRCA1 and RAD51 in G1. f, Quantification of fold change of RPA, BRCA1 and RAD51 recruitment at centromeric DSBs in siSETD1A vs siSCR NIH3T3 cells in G2 and expressing Cas9 + mi gRNA. g, Quantification of fold change of RPA, BRCA1 and RAD51 recruitment at centromeric DSBs in NIH3T3 cells in G2, expressing Cas9 + mi gRNA and co-expressing dCas9-LSD1 or dCas9 alone. h, Western blot analysis of flag-SETD1A and flag-SETD1A-ΔSET in NIH3T3 cells using flag antibody. i, Chromatin immunoprecipitation analysis of H3K4me2 enrichment at centromeres over the Input and relative to the no-antibody control in cells depleted for SETD1A (siSETD1A) and reconstituted with WT SETD1A or with a truncated catalytically inactive mutant (SETD1A-ΔSET) or in control cells (siSCR). j, Quantification of fold change of RAD51 recruitment in G2 at pericentromeric DSBs in NIH3T3 cells expressing Cas9 + mi gRNA and siSETD1A relative to siSCR. For definition of p values, statistic method and sample number see Statistics and Reproducibility section.

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