T-cell potential of Sca-1⁺c-Kit⁺ cells generated by culturing with Delta1^ext-IgG. (A) Cultured LSK (Ly5.2) cells were stained for Sca-1 and c-Kit and sorted into a fraction containing Sca-1⁺ c-Kit⁺ cells and another fraction containing the rest of the population (Sca-1⁻ cKit⁺, Sca-1⁺c-kit⁻, and Sca-1⁻c-kit⁻) and injected into lethally irradiated C57BL/6 mice with 1 × 10⁵ Ly5.1-competing BM cells. (B) CBCs and FACS analysis were performed on blood samples at 3 weeks after HCT to determine the number of CD3⁺ cells generated in mice receiving either Sca-1⁺ c-Kit⁺ cells (○) or Sca-1⁻cKit⁺, Sca-1⁺c-kit⁻, and Sca-1⁻c-kit⁻ (▿). Data represent the mean plus or minus SEM. (C) Cultured cells were gated for Sca-1– and c-kit–expressing cells and sorted according to the presence or absence of B220, CD25, Thy1, or Il7Rα expression. (D) Lethally irradiated C57BL/6 recipients received 1 × 10⁶ Ly5.2 SK-lymphoid–positive cells and 1 × 10⁵ Ly5.1 BM cells (closed bar) or 1 × 10⁴ Ly5.2 SK-lymphoid–negative cells and 1 × 10⁵ Ly5.1 BM cells (open bar). FACs analysis and CBCs were performed on blood samples obtained at various times after HCT. At 5 weeks, lymphoid (CD3, CD4, CD8, and CD19) and myeloid (GR1 and F4/80) engraftment were compared between the 2 groups. Data represent the mean plus or minus SEM.