Fig. 5
From: BRCA2 controls DNA:RNA hybrid level at DSBs by mediating RNase H2 recruitment
RNase H2 is recruited to DSBs preferentially in the S/G2-phase of the cell cycle. a ChIP of RNASEH2A (top) and γH2AX (bottom) at the AsiSI cut site in uncut or cut DIvA cells. The bar graph shows the fold induction in cut cells (6 h after AsiSI induction) compared to uncut. Error bars represent s.e.m. (n ≥ 2 biological replicates). b Representative images of PLA between RNASEH2A and γH2AX in not irradiated (no ir) or irradiated (2 Gy) U2OS cells. Scale bar: 10 μm. c Dot plot shows the number of signals per nucleus of PLA between RNASEH2A and γH2AX in not irradiated (no ir) or irradiated (2 Gy) U2OS cells. At least n = 200 cells were counted from four independent experiments. Lines represent mean ± s.e.m. d Representative images of PLA between RNASEH2A and γH2AX in not irradiated (no ir) or irradiated (2 Gy) HeLa-FUCCI cells. Scale bar: 10 μm. e Dot plot shows number of signals per nucleus of PLA between RNASEH2A and γH2AX in not irradiated (no ir) or irradiated (2 Gy) HeLa-FUCCI cells. At least n = 170 cells were counted from three independent experiments. Lines represent mean ± s.e.m. f Representative pictures of super-resolution imaging analysis of γH2AX (cyan) and RNASEH2A (magenta) colocalization in G1- or S-phase synchronized NCS-treated U2OS cells. Scale bar: 5 μm. g Dot plot shows the normalized number of overlaps relative to random of γH2AX and RNASEH2A signals in G1- or S-phase cells. At least n = 50 events were counted from three independent experiments. Lines represent mean ± s.e.m. **P < 0.01, ****P < 0.0001 (two-tailed Student’s t test). Source data are provided as a Source Data file
