Extended Data Fig. 1: Enforced Trmt6/61a mildly disturbs the homeostasis of hematopoietic system at young age.

(a) The schematic diagram showing the strategy to generate SFFV-Trmt6/61a-transgenic mice. (b-c) 3-month old Trmt6/61a^TG or WT mice were analyzed for white blood cell (WBC), lymphocyte (LYM), neutrophil (NEUT), red blood cell (RBC), platelet (PLT), myeloid, B cells, and T cells. (b) The scatter plots show the count of WBC, LYM, NEUT, RBC and PLT between Trmt6/61a^TG and WT mice. (c) The scatter plots depict the percentage of B, CD4⁺ T, CD8⁺ T cells, and myeloid in peripheral blood. n = 12 mice per group. (d-h) 8⁻week-old Trmt6/61a^TG and age-matched WT mice were analyzed for analyzed for myeloid, B cells, T cells, progenitors and HSCs. (d) Percentage of myeloid, B, CD4⁺ T and CD8⁺ T cells in bone marrow. (e and f) This histogram shows the frequency (e) and the number per femur (f) of CMP, GMP, MEP, and CLP between Trmt6/61a^TG and WT mice. (g and h) This histogram shows the frequency (g) and the number per femur (h) of LT-HSC, ST-HSC and MPP between Trmt6/61a^TG and WT mice. The surface marker combinations are listed in Supplementary Table 3. n = 8 WT mice and n = 6 Trmt6/61a^TG mice. (i-m) 30 HSCs from either 2-month old Trmt6/61a^TG mice or age-matched WT mice were transplanted into lethally irradiated recipients together with 2×10⁵ competitor cells. Chimera in peripheral blood and bone marrow was checked every month until the 3^rd month. (i) Percentage of donor cell reconstitution in overall (CD45.2⁺), B (B220⁺), T (CD3⁺) and myeloid (Mac-1⁺) cell every month after transplantation. (j and l) The lineage distribution in donor-derived peripheral blood (j) and bone marrow (l) at the 3^rd month. (k and m) Percentage of donor cell reconstitution in bone marrow (k) and HSC (m). n = 9-10 mice per group (WT: n = 9; Trmt6/61a^TG: n = 10). Two-tailed unpaired Student’s t-test was used for statistical analysis in b and i. Data are shown as the mean ± SD in c-m.

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