Ebf-1 is critical for IL-7 mediated lineage restriction in the CLP compartment. (A) Diagrams with the overall cloning frequency (black bars) and the development of CD19⁺ (gray bars), mixed lineages (CD19⁺ cells and NK1.1⁺ cells (striped bars), or NK1.1⁺ (white bars) from single wt (Ebf-1^+/+) Ebf-1 heterozygote (Ebf-1^+/−) or Ebf-1–deficient (Ebf-1^−/−) CLPs as indicated. The data are collected from 3 independent experiments using 5- to 6-week-old Ebf-1–deficient mice, and the data are mean + SEM. (B) Representative FACS analysis displaying LY6D⁺ and LY6D⁻ populations within the Flt-3⁺IL-7R⁺Sca1^lowKit^low CLP from BM (femurs and tibias) of normal (Ebf-1^+/+) and Ebf-1–deficient (Ebf-1^−/−) mice. The numbers in the plots are mean percentages of LY6D^+/− cells of total CLP analyzed in 2 independent experiments. (C) Diagrams with the percentage of CD19⁺, NK1.1⁺, or CD11c⁺ cells obtained in culture after seeding single Ly6D⁺ CLPs from wt (Ebf-1^+/+) or Ebf-1–deficient (Ebf-1^−/−) mice. Each dot represents a single clone. The difference between 2 genotypes was evaluated by 2-tailed t test. The graph in panel D displays the cloning frequency of wt and Ebf-1–deficient Ly6D⁺ CLPs after 5 days of incubation under myeloid conditions. The data are collected from 288 seeded Ebf-1–deficient and 462 wt cells collected from 2 independent experiments. The error bars indicate SD.