Extended Data Fig. 2: Zrsr2 loss enhances self-renewal of hematopoietic stem cells (HSCs).

a, FACS of cells from 5th methylcellulose plating (see Fig. 1b) for live, LSK cells. Wild-type bone marrow (BM) from 6-week old mouse (left) as a staining control. b, Number of CFU-GM, CFU-GEMM, and BFU-E colonies from initial plating of LT-HSCs (lin^- LSK CD150⁺ CD48^-) from Mx1-cre control, Mx1-cre Zrsr2^fl/y, and Mx1-cre Zrsr2^fl/fl mice into methylcellulose. Mean ± SD, two-sided t-test. n = 4 biologically independent experiments. c, Schematic of competitive BM transplantation. d, Number of methylcellulose colonies from LT-HSCs from Mx1-cre control, Mx1-cre Zrsr2^fl/x, Mx1-cre Zrsr2^fl/y, and Mx1-cre Zrsr2^fl/fl. Mean value ± SD. n = 3 biologically independent experiments. e, Box-and-whisker plots of percentage of peripheral blood CD45.2⁺ cells in competitive transplantation pre- and post-pIpC using CD45.2⁺ Mx1-cre control, Mx1-cre Zrsr2^fl/x, Mx1-cre Zrsr2^fl/y, and Mx1-cre Zrsr2^fl/fl mice. For box and whiskers plots throughout, bar indicates median, box edges first and third quartile values, and whisker edges minimum and maximum values, two-sided t-test. Experiment repeated three times with similar results in (d) and (e). f, Percentage of CD45.2⁺ B220⁺ (left), CD45.2⁺ CD11b⁺ Gr1⁻ (middle), and CD45.2⁺ CD3⁺ cells (right) in primary competitive transplantation. Mean ± SD. P values by two-sided t-test using the values at 16 weeks after transplant. P value relative to the control group at 16 weeks by a two-sided t-test. g, FACS analysis and gating strategy of BM cells from representative primary recipient mice in competitive transplantation. h, Box-and-whisker plots of numbers of ST-HSCs, CMPs, MEPs, and pDCs in BM of primary recipient mice in competitive transplantation. For box and whiskers plots throughout, bar indicates median, box edges first and third quartile values, and whisker edges minimum and maximum values. P value relative to control by a two-sided t-test.

Source data