Kinetics of CLP^F-driven T cell reconstitution. (A) Kinetic analyses of B-cell (squares) and T-cell (diamonds) reconstitution of spleen (left panel) and blood (right panel) of sublethally irradiated mice that received transplants of 10³ CLP^F (black) or Flk2⁻ fraction cells (white). Results shown are mean values of 3 to 5 animals analyzed per time point. For blood analysis, the same animals were bled periodically for 10 weeks. (B) FACS plots illustrating the CD4/CD8 profile of host (top row) and CLP^F derived donor cells (bottom row) as well as reconstitution with thymic B cells (B220⁺), γδ T cells, and NK cells showing the lymphoid potential of CLP^Fs. (C) FACS plots illustrating the T-lineage development of intravenously injected CLP^F in the thymus, from 1 to 5 weeks. Detailed analysis of CD4/CD8 development (top row), and within the CD4/CD8 double negative fraction, CD44 and CD25 development (middle row) or CD25 and c-Kit development (bottom row), after previously gating on lineage negative cells. Shown are representative FACS plots from 3 mice that received transplants per time point. (D) Thymic reconstitution 6 days after intrathymic (i.t.) injection transfer of 10³ CLP^F into congenic unirradiated mice. Events shown are gated on lineage negative donor-derived cells. FACS analysis reveals generation of c-Kit^hi DN1 (CD25⁻c-Kit⁺) and c-Kit^hi DN2(CD25⁺c-Kit⁺) directly derived from CLP^F. Numbers on plots are percentages of gated cells.