Fig. 5

Rare EDC4 germline variants identified in breast cancer patients are functionally deleterious. a Pedigrees of the five breast cancer families with EDC4 mutations (based on NP_055144). The probands are indicated by arrows, and slashed symbols denote deceased individuals. The ages at diagnosis and/or death are included when known. Individuals genotyped for mutations are marked either as carriers (+) or non-carriers (−). Additional clinical annotations are depicted as shown in the inset. b Diagram showing that all the residues mutated in BRCA1/2-mutation-negative patients cluster inside or close to the WD40 domain of EDC4. c Functional studies of mutations found in patients shown in a. All EDC4 mutants fail to revert the DEB sensitivity phenotype of HEK293T EDC4−/− cells. Graph shows survival after DEB treatment of five EDC4 mutants generated in HEK293T cells. Data shown represent results from at least two combined independent experiments. Error bars indicate mean ± s.d. Means were statistically compared using the two-tailed Student’s t test: EDC4−/− (p = 0.000; p = 0.000), EDC4 corrected (p = 0.760; p = 0.748), G42E (p = 0.002; p = 0.005), S277R (p = 0.024; p = 0.011), D361E (p = 0.001; p = 0.001), R471Q (p = 0.010; p = 0.017), and V477M (p = 0.000; p = 0.000). d HEK293T EDC4−/− cells expressing the mutants show increased G2/M block induced by DEB treatment. e HEK293T EDC4−/− cells expressing the mutants show DEB-induced chromosome fragility as shown with the flow cytometric MN assay. Data shown in d, e represent results from at least three combined independent experiments. Error bars indicate mean ± s.d. Means were statistically compared using one-way analysis of variance (ANOVA) followed by a Dunnett’s multiple comparison test