Fig. 4
BRCA2 suppresses replication stress associated with G1 53BP1 nuclear bodies. a BRCA2 ∆Ex3-4/– cells were serum starved with or without release, as in the schematic (top), and γH2AX mean nuclear intensities were quantified (bottom). γH2AX mean nuclear intensities are shown from one experiment, which is representative of two independent experiments. Median γH2AX intensity, red bars. b 53BP1 nuclear body analysis in G1 phase. Cyclin A–nuclei (indicating G1 phase) are outlined (left). Quantification of 53BP1 nuclear body distribution is shown (right). n = 4. Scale bars 10 µm. c S phase DNA damage analysis. Where indicated, cells were treated with EdU for 30 min followed by HU treatment (4 mM, 5 h) with or without release (Rel.) before harvest. Quantification of γH2AX mean nuclear intensities is shown from one experiment, which is representative of two independent experiments. Box and whiskers show the 10th and 90th percentiles. d HU-induced 53BP1 nuclear body formation analysis, as in the schematic (top). Sample plots for high content image cytometry analysis are shown on the left. The EdU+ G1 cell fraction (i.e., EdU+, cyclin A–, 1N DNA content) for 53BP1 nuclear body quantification is highlighted. Quantification of 53BP1 nuclear body distribution in EdU+ cells in G1 at the time of harvest is shown in the graph. n ≥ 3. Error bars s.d. ns, not significant; ****p < 0.0001 (two-tailed Mann–Whitney test)
