Extended Data Fig. 6: Loss of SIRT2 alters T cell development under competitive bone marrow reconstitution.

a–c, representative flow-cytometric plots showing CD4 and CD8 distribution in WT and Sirt2^−/− thymi from C57BL/6 (a), OT-II (b), and PMEL (c) mice. Frequencies of DN (DN1–DN4), DP and SP thymocytes for CD4⁺ and CD8⁺ subsets are indicated in the right panel. Each dot = one mouse (n = 3 per group). d, schema of experimental design of mixed bone marrow chimeras generated by reconstituting lethally irradiated C57BL/6 × C57BL/6.SJL F1 recipient mice (n = 4) with a 1:1 mixture of bone marrow cells from WT (CD45.1⁺) and Sirt2⁻/⁻ (CD45.2⁺) donors. Mice were sacrificed 8 weeks after bone marrow transplantation. e, Flow-cytometric plots of WT CD45.1⁺ and Sirt2^−/− CD45.2⁺ bone marrow cells from WT (CD45.1), Sirt2^−/− (CD45.2) donor mice and the 1:1 mix prior injection. f, Flow-cytometric plots of hematopoietic stem cells (HSCs) identified as lineage⁻ (Lin⁻), mature T cell⁻ (CD3⁻), c-kit⁺ and Sca-1⁺ (left) and the distribution of WT CD45.1⁺ and Sirt2^−/− CD45.2⁺ HSC at 8 weeks post-reconstitution in the bone marrow of recipient mice. g-i, Flow-cytometric plots of donor-derived WT CD45.1⁺ and Sirt2^−/− CD45.2⁺ DN1–DN4, DP, CD4⁺ SP and CD8⁺ SP thymocytes in the thymus of lethally irradiated C57BL/6 × C57BL/6.SJL F1 recipient mice analyzed 8 weeks after reconstitution with a 1:1 mixture of WT and Sirt2^−/− bone marrow cells. Analysis was based on the distribution of CD4 and CD8 markers in CD45.1⁺ (WT) and CD45.2⁺ (Sirt2^−/−) T cells. DN stages (DN1–DN4) were defined by CD44 and CD25 expression, and DN1 cells were further assessed for c-Kit⁺Sca-1⁺ expression. j,k Flow-cytometric plots of WT CD45.1⁺ versus Sirt2^−/− CD45.1⁺ CD4⁺ and CD8⁺ T cells in the spleen (j) and lymph nodes (k) of recipient mice as in d at 8 weeks post-reconstitution. Data are mean ± SEM. P values are determined by two-way ANOVA (a-c). Data are representative of three (a-c) and four (d-k) independent experiments.

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