Figure 4: COBRA1 is not involved in BRCA1-mediated HR. | Nature Communications

Figure 4: COBRA1 is not involved in BRCA1-mediated HR.

From: Genetic suppression reveals DNA repair-independent antagonism between BRCA1 and COBRA1 in mammary gland development

Figure 4

(a) Diagram of the GFP reporter assay for measuring HR efficiency. I-Scel: restriction enzyme. iGFP: internal GFP fragment as the template for HR. (b) Immunoblot of COBRA1 and BRCA1 for assessing siRNA knockdown efficiency with control (Con) oligos or ones targeting human BRCA1 (-B) and COBRA1 (-C) in HeLa cells. (c) Percentage of GFP+ cells as a result of HR-mediated DSB repair. Results are average of three independent experiments, and were normalized to the control group. *P<0.05 by Student’s t-test. (d) Mice of 8-wk old were pulse-labelled with BrdU, irradiated (20 Gy), and mammary glands were harvested 3 h later for immunostaining for γH2AX and BrdU. Scale bar, 5 μm. (e) The same samples as shown in d were stained for Rad51 and BrdU. Scale bar, 5 μm. (f) Percentage of Rad51+/BrdU+ mammary epithelial cells. *P<0.05 by Student’s t-test. The numbers of animals used are indicated below the graph. Error bars represent s.e.m. wk, week.

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