Functional evaluation of BRCA2 variants located at the C-terminal domain. (A) Chart summarizing the sensitivity of ES cells expressing different mutant BRCA2 to different DNA-damaging agents. (B) Quantification of RAD51 and γ-H2AX foci after ionizing radiation. Error bars represent the mean ± SD. (C) HR efficiency as measured by gene targeting to the Rosa26 locus. For each variant and WT BRCA2, 2 independent clones were used, and they are marked 1 and 2. (D) Total number of chromosomal abnormalities present in WT and different mutant ES cells are represented. Randomly selected 100 metaphase spreads were counted blindly in each case. Two independent clones were used for each mutant and WT cell. (E) Representative metaphase spreads are shown. Thick arrows indicate different chromosomal abnormalities. To rule out the possibility of secondary mutations, each experiment was conducted using at least 2 independent clones of each mutant and they behaved similarly.