Figure 3.

MDP and MP are progenitors of only cDC-like cells in co-cultures. (A) BM from CX3CR1-GFP mice was stained with antibodies and analyzed flow cytometrically for sorting MDP and MP as described in Figure 1. Lin⁻ cells were distinguished on a forward scatter (FSC) plot and c-kit^hi cells gated to distinguish Flt3⁺ cells. These were then sorted as CX3CR1⁺ and CX3CR1⁻ subsets of MP and MDP, respectively, each of which showed CD115 staining. (B) Sorted cells were co-cultured over 5G3 stroma using Lin⁻ BM as a control. Non-adherent cells were collected from co-cultures at different time points and stained for CD11c, CD11b, MHC-II, and CD8α to detect production of CD11b⁺CD11c⁺CD8α⁻B220⁻MHC-II⁻ L-DC and CD11b⁺CD11c⁺CD8α⁻B220⁻MHC-II⁺ cDC-like cells, and their expression of CX3CR1. Data shown reflect cells produced at 21 days. (C) Cell production was calculated in terms of number of cells of each type produced. Graphs show mean ± SE for triplicate co-cultures.