Figure 1. Dnmt3a^R878H/+ expands HSCs and multipotent progenitor cells.

(A) Schematic diagram of the design of the Dnmt3a^fl-R878H/+ allele. * indicates 2633G>A in exon 23 to encode the R878H mutation. (B) Frequency and (C) Total number of LTHSC, STHSC, MPP2, MPP3, MPP4 and MyPro cells in the BM of +/+ (n = 4) and R878H/+ (n = 6) mice at 6 months post-pIpC. (D) Total colony-forming units (CFU) and (E) Colony types derived from 50K BM MNCs isolated from +/+ (n = 6) and R878H/+ (n = 8) mice at 4 months post-pIpC. G (granulocyte), M (macrophage), GM (granulocyte-macrophage), BFUE (burst forming-unit erythroid), GEMM (mixed granulocyte-erythroid-macrophage-megakaryocyte). Results are from 3 independent experiments. (F) Experimental design for competitive transplantation of +/+ or fl-R878H/+ BM cells with wild-type B6.CD45.1 BM cells at a 1:1 ratio into lethally irradiated B6.CD45.1 recipient mice followed by pIpC to induce recombination of Dnmt3a^fl-R878H. (G) Frequency of donor-derived (CD45.2⁺) +/+ (n = 9) or R878H/+ (n = 9) cells in PB of recipient mice post-pIpC. Results are from 2 independent experiments. (H) Frequency of myeloid (CD11b⁺), B (B220⁺) and T (CD3⁺) cells within donor-derived (CD45.2⁺) PB and (I) Gr-1⁺ and Gr-1⁻ cells within donor-derived myeloid PB at 6 months post-pIpC. (J) Frequency of LTHSC, STHSC, MPP2, MPP3 and MyPro cells within donor-derived +/+ (n = 5) or R878H/+ (n = 5) BM at 6 months post-pIpC. Results are from 2 independent experiments. In all graphs, dots represent individual mice, bars indicate mean ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001.