Progenitors are enriched within the c-kit⁺CD34⁺ subset of neonatal spleen. (A) Eight-day-old splenocytes were sorted to give c-kit⁺CD34⁻ and c-kit⁺CD34⁺ populations (Expt B). These were cultured over STX3 to assess DC development. (B) Over time, nonadherent cells were collected from cocultures and assessed for DC production flow cytometrically. Cells were simultaneously stained for CD11c, CD11b, CD8α, and MHC-II. FSC and SSC analysis was used to gate large cells, and isotype controls were used to indicate background antibody binding (density plots) and set gates. Square and round gates identify common populations across different plots, and percentage of cells showing specific binding is indicated within gates. Staining for CD8 was negative (data not shown). Results are reflective of 2 similar experiments.