Fig. 3: Effects of PALB2 VUSs on G2/M checkpoint response.

a Detection of the PALB2 nonsense variant (c.1802T > A, p.Y551*) of the EUFA1341 cell line. EUFA1341 cells express truncated PALB2 proteins lacking the ability to recruit BRCA2-RAD51. b Western blotting analysis of FLAG-tagged PALB2 expression in the EUFA1341 stable cell lines. c G2/M checkpoint response in EUFA1341 cells reconstituted with the empty vector (EV) or wild-type (WT) PALB2. Cells were treated with 3 Gy of IR and collected at indicated time points to measure the mitotic index by flow cytometry. Data represent the mean percentage (±SEM) of mitotic cells relative to the untreated cells (NT) from three independent experiments. d Representative flow cytometric plots of mitotic cells by phospho-histone H3 (Ser10) and propidium iodide (PI) staining. e G2/M checkpoint response in EUFA1341 cells stably expressing PALB2 constructs (or an empty vector, EV). Cells were analyzed before and at indicated time points after 3 Gy of IR. Data represent the mean percentage (±SEM) of mitotic cells relative to the untreated cells (NT) from three independent experiments. Statistical significance was analyzed by one-way ANOVA and Dunnett’s multiple comparisons test. The values of cells expressing indicated constructs were compared to cells expressing the WT PALB2 protein at the same time point. *P < 0.05; **P < 0.01; ***P < 0.001.