Fig. 4: The hypomorphic missense p.(Glu2599Gly) variant has partial HRR activity.

a Only the BRCA2 p.(Glu2599Gly) variant is expressed. Immunoblotting of BRCA2 in wt and BRCA2 KO HEK293T transiently transfected with pCDNA3 wt BRCA2, empty vector (EV), BRCA2 p.(Ile605Asnfs*11) and BRCA2 p.(Glu2599Gly), 48 h after transfection. Vinculin antibody was used as a loading control on the same sample run and processed in parallel. b BRCA2 p.Glu2599Gly variant shows partial complementation of RAD51 foci formation. IR-IF analysis of RAD51 in HEK293T BRCA2 KO cell line expressing wt BRCA2, EV, BRCA2 p.(Ile605Asnfs*11) and BRCA2 p.(Glu2599Gly), 6 h after 5 Gy. Mean of three independent experiments represented. Error bars show s.d. Statistics are done with the One-Way ANOVA test and Bonferroni correction **P < 0.01, ***P < 0.001. c Intermediate HR efficiency of p.(Glu2599Gly) BRCA2 expressing cells. GFP expression from the DR-GFP reporter was analysed as a measure of HR activity. The percentage of GFP-positive cells for each variant was shown. Results represent the mean of at least four independent experiments. Error bars represent s.d. Statistics are done with the One-Way ANOVA test and Newman–Keuls multiple comparison tests, *P < 0.05, **P < 0.01, ***P < 0.001.