Fig. 2
From: BRCA2 controls DNA:RNA hybrid level at DSBs by mediating RNase H2 recruitment
DSB-induced DNA:RNA hybrids form at resected DNA ends in S/G2-phase cells. a DRIP-qPCR analysis at 1.5 kb on the right from the I-PpoI cut site within DAB1 gene in G1- or S/G2-phase-sorted HeLa-FUCCI cells transfected with the I-PpoI nuclease. The bar graph shows fold induction of DNA:RNA hybrid levels in cut samples relative to uncut. Error bars represent s.e.m. (n = 3 independent experiments). b Representative pictures of super-resolution imaging analysis of γH2AX (cyan) and DNA:RNA hybrids (yellow) colocalization in S-phase synchronized U2OS cells treated with neocarzinostatin (NCS). Scale bar: 5 μm. c Dot plot shows the normalized number of overlaps relative to random of γH2AX and DNA:RNA hybrids signals in G1- or S-phase NCS-treated U2OS cells. At least n = 60 events were counted from three independent experiments. Lines represent mean ± s.e.m. d DRIP-qPCR analysis at 1.5 kb on the right from the I-PpoI cut site within DAB1 gene in cells knocked-down for EXO1. Error bars represent s.e.m. (n = 3 independent experiments). e Strand-specific RT–qPCR analysis of dilncRNAs levels at 1.5 kb on the right from the I-PpoI cut site within DAB1 gene in cells knocked-down for EXO1. Error bars represent s.e.m. (n = 4 independent experiments). f DRIP-qPCR analysis at 1.5 kb on the right from the I-PpoI cut site within DAB1 gene in cells knocked-down for CtIP at different time points after cut induction. Error bars represent s.e.m. (n = 5 independent experiments). g Strand-specific RT-qPCR analysis of dilncRNAs levels at 1.5 kb on the right from the I-PpoI cut site within DAB1 gene in cells knocked-down for CtIP at different time points after cut induction (n = 2 independent experiments). *P < 0.05, **P < 0.01, ****P < 0.0001 (two-tailed Student’s t test). Source data are provided as a Source Data file
