Extended Data Fig. 9: Loss of intestinal epithelial integrity contributes to expansion of pre-leukaemic cells via ALPK1.

a, Experimental design to examine the effect of gut injury. BM cells from three Dnmt3a^f/f;Mx1-Cre mice or three Dnmt3a^f/f;Alpk1^KO;Mx1-Cre mice treated with poly(IC) to induce recombination were transplanted into WT mice (n = 20 mice per group) to generate chimeric mice. Chimeric mice were treated with either water (H₂O) or DSS (2.5%) for 1 week, and flow cytometry was performed on BM. Secondary transplants were performed with purified donor HSCs (CD45.2+). b, Number of donor HSCs (Lin-cKit+Sca1 + CD150 + CD48-) in the BM (n = 10 mice per group). c, Donor-derived proportions in PB at the indicated time points (n = 8 mice per group). d, H&E-stained BM, spleen, and liver isolated from Alpk1^WT (upper panel) and Alpk1^KO (bottom panel) mice. Images are representative of four replicates. e, Complete blood counts of Alpk1^WT and Alpk1^KO mice (n = 8 mice per group). WBC, white blood cells; NE, neutrophils; LYM, lymphocytes; RBC, red blood cells; Hb, haemoglobin; PLT, platelets. f, Outline of competitive BM transplantation experiment where BM cells from Alpk1^WT or Alpk1^KO mice (n = 1 mouse per group), and BM cells from one WT mouse (CD45.1) were transplanted competitively (1:1) into WT mice (CD45.1; total 1×10⁶ cells per mouse) (n = 6 mice per group). g, Donor-derived proportions in PB at the indicated time points (n = 6 mice per group). h, Donor-derived multilineages, B (CD19+ B220+), T (CD3+), and myeloid (Gr1+ CD11b+) cells in the PB (n = 6 mice per group). Error bars represent the SEM. *, P < 0.05, **, P < 0.01, ***, P < 0.001. P values were calculated for the indicated comparisons using a two-tailed unpaired Student’s t-test (b) and a two-way ANOVA (c, e, g, and h).

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