Further characterization of precursors in neonatal spleen. (A) Eight-day-old splenocytes were sorted into 3 populations of c-kit⁻, c-kit^lo, c-kit^hi cells (Expt A) and cocultured in equal number over STX3. (B) Cell production was monitored by photography under phase microscopy after 13 days. Bar represents 50 μm. (C) Nonadherent cells were collected from individual cocultures at 13, 23, and 30 days. Cells were simultaneously stained for CD11c, CD11b, CD8α, and MHC-II. FSC and SSC analysis was used to gate large cells for marker analysis to assess DC production. Isotype controls were used to determine background binding (density plots) and set gates. Square and round gates were used to identify common populations across different plots. Percentage of cells showing specific staining is indicated within gates. Staining for CD8 was negative (data not shown). Results are reflective of 2 similar experiments.