Fig. 2: Hematological analysis of young polygenic Brca2^Δ27/Δ27 + Rad51c^dah/dah mutant mice reveals pre-state of bone-marrow failure.

a Bar graph of nucleated cells per femurs of mice between 8–12 weeks. Data represent biologically independent samples from wild-type (n = 7), Brca2^Δ27/Δ27(n = 6), Rad51c^dah/dah (n = 6) and Brca2^Δ27/Δ27 + Rad51c^dah/dah (n = 8) mice. Brca2^Δ27/Δ27 + Rad51c^dah/dah against WT: p = 0.0099; Brca2^Δ27/Δ27 + Rad51c^dah/dah against Rad51c^dah/dah: p = 0.0106. b Schematic of hematological cell lineages. HSC, hematopoietic stem cell. Mye, myeloid progenitor. CLP, common lymphoid progenitor. Pl, platelets. Ery, Erythrocytes, red blood cells. Leu, Leukocytes, white blood cells. Gran, Granulocytes. Eos, Eosinophils. Bas, Basophiles. Neutr, Neutrophils. Mono, Monocytes. Macro, Macrophages. Lym, Lymphocytes. T, T-cells. B, B-cells. NK, natural killer cells. c–e Complete blood count analysis of 8–12 weeks old Brca2^Δ27/Δ27 + Rad51c^dah/dah and control mice. Data represent biologically independent samples from wild-type (n = 7), Brca2^Δ27/Δ27(n = 5), Rad51c^dah/dah (n = 11) and Brca2^Δ27/Δ27 + Rad51c^dah/dah (n = 7) mice. c Peripheral white cell concentration. d Peripheral red cell concentration. Brca2^Δ27/Δ27 + Rad51c^dah/dah against Rad51c^dah/dah: p = 0.0271. e Mean corpuscular volume as a measure for macrocytosis. Brca2^Δ27/Δ27 + Rad51c^dah/dah against WT: p = 0.0321; Brca2^Δ27/Δ27 + Rad51c^dah/dah against Rad51c^dah/dah: p = 0.0005. f–i Hematological lineage analysis by flow cytometry was performed for bone marrow cells collected from 8–12-weeks-old mice with indicated genotypes. f Bar graph of common myeloid progenitor population. Data represent 4 biologically independent samples for each genotype. g Bar graph of common lymphoid progenitor population. Data represent 4 biologically independent samples for each genotype. h Bar graph of hematopoietic stem cell population. Data represent biologically independent samples from wild-type (n = 5), Brca2^Δ27/Δ27(n = 4), Rad51c^dah/dah (n = 5) and Brca2^Δ27/Δ27 + Rad51c^dah/dah (n = 6) mice. i Bar graph of colony formation capacity (CFU) of bone marrow progenitor cells per 30.000 plated cells. Data represent independent samples for wild-type (n = 8), Brca2^Δ27/Δ27(n = 5), Rad51c^dah/dah (n = 6) and Brca2^Δ27/Δ27 + Rad51c^dah/dah (n = 13). Brca2^Δ27/Δ27 + Rad51c^dah/dah against WT: p < 0.0001; Brca2^Δ27/Δ27 + Rad51c^dah/dah against Rad51c^dah/dah: p = 0.0002. j Schematic of 16-week repopulation assay workflow. Briefly, bone marrow cells from female wild-type (WT) and male mutant mice were mixed and allowed to repopulate in female mice that were depleted for their bone-marrow. After 16 weeks, the repopulation capacity of mutant bone marrow is measured by PCR of male cells in the blood, which were contributed by the mutant mouse. k Quantitation of repopulation assay. Error bars represent the standard error of the mean. Red bar in scatter graphs represent the mean. Data represent biologically independent animals with Brca2^Δ27/Δ27(n = 3) and Brca2^Δ27/Δ27 + Rad51c^dah/dah (n = 4) genotypes. Brca2^Δ27/Δ27 against Rad51c^dah/dah: p = 0.0006. p-values (A-I) are derived using the one-way ANOVA test. * p < 0.05, *** p < 0.001, **** p < 0.0001. The p-values (K) are derived using the two-tailed Student t test. *** p < 0.001.