Extended Data Fig. 1: Characterisation of Dnmt3aR882H/+ mouse model.
From: Mitochondrial metabolism sustains DNMT3A-R882-mutant clonal haematopoiesis
(a) Schematic representation of genotyping (red arrows) and recombination primers (blue arrows) binding to WT, Dnmt3aflox-R882H and Dnmt3aR882H alleles. (b) Genotyping confirmed the presence of the Dnmt3aR882H allele pre-pIpC in both Dnmt3aR882H/+ and Dnmt3aR882H/+ Cre genotypes (top panel, left half). After Cre induction with pIpC the Dnmt3aR882H allele completely disappears from Dnmt3aR882H/+ Cre mice, indicating complete recombination in BM (top panel, write half). Mid panel illustrates the presence of the Mx1-Cre allele in Dnmt3aR882H/+ Cre genotypes. Lower panel shows recombination in Dnmt3aR882H/+, Cre mice post-pIpC. A low level of recombination is also observed in Dnmt3aR882H/+, Cre mice pre-pIpC (lower panel, left half). However, the full recombination was not observed pre-pIpC Dnmt3aR882H/+ Cre, indicated by the presence of Dnmt3aR882H allele (top panel, left half). (c) Frequency of LT-HSC, (d) Lin-ve, (e) ST-HSC, (f) LSK, MPP, LMPP and (g) CLP progenitors in BM of Dnmt3aR882H/+ and WT mice 6-weeks post-pIpC, n = 6 mice per group in c-g (h) Proportion of B, T and myeloid cells in BM, n = 5 mice per group and (i) Sp of Dnmt3aR882H/+ and WT mice 6-weeks post-pIpC, n = 6 mice per group. The mean ± SEM is shown in c-i (j) Proportion of myeloid, B and T cells in PB, n = 4 mice per group. (k) Frequencies of LK, GMP, CMP, MEP, n = 4 mice for WT and n = 3 mice for Dnmt3aR882H/+ (l) Frequencies of LSK, MPP, LMPP (m) LT-HSC, ST-HSC and (n) CLP in BM of Dnmt3aR882H/+ and WT mice 1-year post-pIpC for j-n. The mean ± SD is shown, n = 4 mice per group for (l-n). (o) WBC, (p) RBC, (q) HGB, (r) PLT, (s) HCT of Dnmt3aR882H/+ and WT mice 20 weeks post-pIpC, n = 5 mice per group for (o-s). P in (h, j, k) by two-sided t-test. MPP, multipotent progenitors; LMPP, lymphoid primed multipotent progenitors; ST-HSC, short-term HSC; GMP, granulocyte-monocyte progenitors; CMP, common myeloid progenitors, MEP megakaryocyte-erythroid progenitors; CLP, common lymphoid progenitors; RBC, red blood cells. Schematic in a was created using BioRender (https://biorender.com).
