Fig. 6.

The E2F1 3KR mutation impairs BRG1 and MRN accumulation at a DSB. a Primary wild-type (WT) and E2f1^3KR/3KR (3KR) MEFs were subjected to the I-PpoI ChIP assay and occupancy of BRG1 and total histone H3 was determined by qPCR at mChrom10 locus. b Purified GST-TopBP1 (BRCT1-6) or GST control protein was incubated with whole-cell extract from wild-type (WT) or E2f1^3KR/3KR (3KR) MEFs that were either untreated (−) or treated (+) with IR (10 Gy) and harvested 2 h post-IR. Associated proteins were pulled down using glutathione beads and western blot was performed with the indicated antibodies. c Primary wild-type (WT), E2f1^S29A/S29A (S29A), and E2f1^3KR/3KR (3KR) MEFs were subjected to the I-PpoI ChIP assay and occupancy of NBS1 and Mre11 was determined by qPCR at mChrom10 locus. d Western blot analysis was performed for phospho-ATM (serine 1981), phospho-p53 (serine 15), γH2AX, and GAPDH using whole-cell extracts from primary wild-type (WT) and E2f1^3KR/3KR (3KR) MEFs, before treatment or 1 h and 3 h post-treatment with 250 ng/ml of NCS. All graphs represent average ± SD of three independent experiments (n = 3). P values were calculated by unpaired Student’s t test. **P ≤ 0.01 is highly significant. Source data of b and d are provided as Supplementary Data 5. Raw data of a and c are in Source Data File