Fig. 5: GATA2 bookmarking is required for definitive haematopoiesis.

a Immunohistochemistry images representing E10.5 wild-type (WT/WT), heterozygous (MD/WT) and homozygous (MD/MD) intra-aortic hematopoietic clusters expressing RUNX1 (red) and CD31 (white) in the ventral (V) or dorsal (D) sides of the dorsal aorta (DA). White arrowheads indicate clusters. Scale bars, 150 µm. b Number (#) of intra-aortic hematopoietic clusters per genotype. n(WT/WT) = 3, n(MD/WT) = 2, n(MD/MD) = 2 embryos. Mean is shown. c–e Colony-forming units for E9.5 yolk sac (c), for E10.5 aorta-gonad-mesonephros (AGM), placenta and foetal liver (d) and for E11.5 AGM and placenta (e) cell suspensions. n(WT/WT) = 2-5, n(MD/WT) = 3−4, n(MD/MD) = 3−5, n(Mut/Mut) = 5 embryos. Mean ± SD is represented. Macrophage (M), granulocyte (G), granulocyte/macrophage (GM), erythroid (E) and mixed colonies (Mix) are shown per embryo equivalent (ee). d, e Statistical significance for the total number of colonies was calculated by one-way ANOVA followed by Bonferroni’s multiple comparison test. d, a, p = 0.01; b, p < 0.001, c, p = 0.002. e, a, b, c and d, p < 0.001. f, Percentage of donor chimerism (CD45.2⁺) 4 weeks (w), and 3 and 6 months (m) after transplantation with E11.5 placenta cells. Red line indicates 1% chimerism. n(WT/WT, 4w, 3m) = 5, n(WT/WT, 6m)=4, n(MD/WT, 4w, 3m, 6m) = 8, n(MD/MD, 4w, 3m, 6m) = 8. a, p = 0.011; b, p = 0.016. g, Quantification of donor contribution to myeloid and lymphoid lineages. Only mice with donor chimerism above 1% were considered. n(WT/WT, 4w, 3m, 6m) = 3, n(WT/WT, 6m) = 4, n(MD/WT, 4w, 3m) = 5, n(MD/WT, 6m) = 4. h Percentage of donor chimerism in bone marrow 6 months after transplantation. n(WT/WT) = 4, n(MD/WT) = 8, n(MD/MD) = 8. a, p = 0.023; b, p = 0.028. f, h Statistical significance at 6 months was calculated with Kruskal-Wallis test followed by uncorrected Dunn’s test. i Percentage of donor chimerism 4 weeks, 3 months and 6 months after competitive transplantation (see Supplementary Fig. 7, Methods). n(WT/WT) = 6, n(MD/WT) = 8. a, p = 0,029. j Quantification of donor contribution to blood lineages. n(WT/WT, 4w, 3m, 6m) = 6, n(MD/WT, 4w, 3m, 6m) = 8. k Percentage of donor chimerism in bone marrow 6 m after transplantation. n(WT/WT) = 6, n(MD/WT) = 8. a, p = 0.020. i, k Statistical significance was calculated with two-tailed Mann–Whitney test at 3 months (i) or at 6 months (k). l Proposed model for the role of GATA2 bookmarking for definitive haematopoiesis. HSPC, hematopoietic stem and progenitor cell. Source data are provided as Source Data file.