Extended Data Fig. 8: Flow cytometric analysis of CD69+ human HSPCs before and after xenotransplantation engraftment.

(a) Human CD34⁺ HSPCs were isolated from the indicated tissues including 20-week gestation fetal liver (20 week) with surface CD69 expression analyzed by flow cytometry at initial thawing (with cells having been frozen at the time of isolation) compared to following 48 hours in culture with the indicated cytokines. Mean fluorescence intensities are shown on each graph. (b) Lineage^- CD34⁺ CD38^- human fetal liver HSPCs expressing or not expressing surface CD69 (based on lowest 10^th percentile and highest 10^th percentile expression) were isolated by fluorescence cell sorting, with a representative post-sort purity analysis shown. (c) Bone marrow mononuclear cells were harvested from NOD.Cg-Prkdc^scid Il2rg^tm1Wj/SzJ (NSG) mice 12 weeks after xenotransplantation with human CD34+ CD38- HSPCs. Representative flow cytometric analysis of human cell engraftment is displayed. Percent of parent population values are shown.