In vivo differentiative potential of spleen precursors. Splenocytes of adult B6.SJL mice were depleted of T and B cells and sorted to give CD11c⁻CD11b⁻(Lin⁻) populations of c-kit⁻ and c-kit⁺ cells. Sorted cells (2 × 10⁴) were transferred intravenously into sublethally irradiated (4.5 Gy) C57BL/6J (CD45.2⁺) recipient mice. Negative “nil” control mice were injected with Hanks balanced salt solution only. For analysis of short-term cell development, spleen was collected at 20 or 41 days and stained with antibodies specific for CD45.1, CD11c, CD11b, CD8α, and MHC-II. Before flow cytometry, cells were incubated with PI (1 μg/mL) for gating of live (PI⁻) cells. Spleens showing CD45.1⁺ progeny (> 0.05%) were examined for donor myeloid cells (CD11b⁺CD8⁻), cDCs (CD11c⁺MHC-II⁺), and lymphoid cells (CD11b⁻CD8⁺). Numbers in gates indicate percentage positive cells. In a second similar experiment, Lin⁻c-kit⁻ and Lin⁻c-kit⁺ cells were sorted and transferred into lethally irradiated (9.5 Gy) host mice and analyzed for donor progeny (CD45.1⁺) after 25 weeks. T-/B-cell depleted spleen was analyzed for DC (CD11c⁺) and myeloid cell (CD11b⁺CD11c⁻) progeny. T-/B-cell enriched spleen was analyzed for lymphoid T (CD8⁺) and B (CD19⁺) cells.