Figure 1. Pathological examination of Dnmt2-deficient mice.

1. Giemsa staining of a femoral section from 8-day-old mice showing endochondral ossification defects. Scale bar: 100 μm.
2. Representative femur sections from 8-day-old wild-type and Dnmt2^−/− mice. The staining of perivascular cells (GLS, green) is strongly reduced in Dnmt2^−/− bone marrow. Nuclei are counterstained with DAPI. Scale bar: 100 μm.
3. Quantitative analysis of capillary numbers. GLS-positive capillaries were counted in wild-type (wt) and Dnmt2^−/− (D2-) mice, in three independent high-power fields (HPF) per mouse (n = 7).
4. Quantification of cell numbers in haematopoietic tissues of 8-day-old mice (n = 8).
5. Representative FACS plots of wild-type and Dnmt2^−/− BM stained for Lin⁻ Sca1⁺ and cKit⁺ cells.
6. Quantification of bone marrow LSK cells in wild-type (three independent experiments, each with pooled BM samples from 4 to 6 mice) and Dnmt2^−/− bone marrow (three independent experiments, each with pooled BM samples of 5–9 mice).
7. Restoration of the LSK cell population in adult Dnmt2^−/− mice. Frequencies of LSK cells were normalized to the frequencies of LSK cells from the bone marrow of wild-type littermates at the indicated age (2–3 independent experiments, each with pooled bone marrow samples from 3 to 9 mice).
8. Increased LSK CD34⁻ and reduced LSK CD34⁺ cell populations in 8-day- and 1-year-old Dnmt2^−/− mice compared to wild-type (two independent experiments, each with pooled bone marrow samples from 3–9 mice).

Data information: Data are presented as mean ± SD. Asterisks indicate statistically significant (P < 0.05, t-test) differences. In (G) and (H), asterisks indicate statistically significant (P < 0.05, t-test) differences relative to wild-type (= 1).