Figure 2. Lis1 is cell-autonomously required for adult hematopoietic stem cell self-renewal.

(a) Average frequency of HSCs (KLSFlt3⁻, KLSF) in whole bone marrow from tamoxifen-treated control (Lis1^+/+; Rosa-creER, indicated as WT) and tamoxifen-treated (Lis1^f/f; Rosa-creER (indicated as Lis1^−/−) mice; n=4 for control (WT), n=3 for Lis1^−/−; *P=0.0140. (b) Representative FACS plots of HSCs (KLSF) from WT and Lis1^−/− mice. (c) Repopulation efficiency of Lis1^−/− HSCs. Representative FACS plots shows donor chimerism (CD45.2⁺ cells) in recipients transplanted with HSCs (KLS CD150⁺ CD48⁻) from WT or Lis1^−/− mice. FACS analysis was performed 28 weeks post-transplantation. (d) Average donor chimerism at different times after transplantation (4–5 mice per cohort). WT is shown with solid squares and Lis1^−/− is shown with open squares. (e) Genome wide expression analysis of Lis1-deficient HSC-enriched cells. Heat map of known regulators of stem and progenitor cell activity significantly affected by the loss of Lis1. (f–i) Lis1 chimeras with hematopoietic-specific Lis1 deletion, (f) Donor chimerism prior to tamoxifen (tam) treatment was assessed two months post-transplantation. (WT) indicates control Lis1^+/+; Rosa-creER and (f/f) indicates Lis1^f/f; Rosa-creER transplanted mice (5 mice in each cohort). (g) Frequency of donor-derived KLS cells in chimeric mice post-deletion. ((WT) +tam) indicates mice that received donor cells from Lis1^+/+; Rosa-creER and ((f/f) +tam) indicates mice that received donor cells from Lis1^f/f; Rosa-creER mice; n=3 for each cohort, *P=0.0277. (h–i) Repopulation ability of whole bone marrow (WBM) cells isolated from Lis1 chimera mice. (h) Representative FACS plots show donor chimerism (CD45.2⁺ cells) in recipients that received cells from either control (f/f +vehicle) or (f/f +tam) Lis1 chimeras. (i) Average donor chimerism at 16 weeks post-transplantation (n=3–4 recipients per cohort; *P=0.0369).