Figure 4.
The progenitors of pDCs are within stem cell–containing fraction and early lymphoid progenitors, and are negatively regulated by Notch receptor ligation. (A) Lin-ckitHiSca1+GFP- (stem cell enriched), Lin-ckit+Sca1- GFP-CD34+FcγRLo (myeloid progenitors), Lin-ckitHi Sca-1+GFP+ (early lymphoid progenitors), Lin-ckitLo Sca-1LoGFP+ (prolymphocytes), and B220+CD19- CD11cLoLy6C- (Ly6C- cohort) were isolated from RAG1/GFP knock-in mice. Cells (10 000) of each kind were cultured for 8 days on OP9 stromal cells with Flt3-L. Cells were harvested and stained with CD45R/B220 and CD19. The B220+CD19- population was sorted and restained with CD11c and Ly-6C (i). The yield of CD11cLoLy-6C+ pDCs recovered per input precursor is shown in the bar graph (ii). CMP indicates common myeloid progenitors. (B) GFP expression was analyzed in B220+CD19-CD11cLoLy6C+ pDCs recovered from HSCs, ELPs, and Pro-L cultures. (C) Lin-ckitHi Sca-1+CD27+ ELPs (10 000) from C57BL/6 BM were cultured for 8 days on OP9-GFP or OP9-DL1 stromal cells. The yield of pDCs per input precursor is shown in the bar graph.