Figure 2
Figure 2. In vivo assay for DC generation from C/EBPα-targeted progenitors. (A) FACS analysis of CD45.2+ donor–derived cells, from 5000 transplanted KSL of C/EBPαfl/fl or Mx1–Cre+ C/EBPαfl/fl–transplanted mice, for CDP (Lin–, Flt3+, FcγRII/III–, cKit+, CD115+, CD11c–) from bone marrow, or cDC (CD19–, CD3–, NK–, CD11c+, MHCII+) and pDC (CD19–, CD3–, NK–, CD11c+, B220+) from the spleen show contribution of donor cells to each population. The data shown are representative dot plots from 3 mice per group for 3 independent experiments with similar results. (B) Absolute numbers of total CD45.2+ CD19–, CD3–, NK– CD11c+ DCs were calculated from total splenocytes (left). B220+ subset of cells was determined to be pDC (black), and B220– MHCII+ cells were cDC (white) and plotted accordingly. CMPs were sorted from either MxCre– C/EBPαfl/fl or MxCre+ C/EBPαfl/fl mice and analyzed for expression of CX3CR1 by RT-PCR. The percentage of CDP from total CD45.2+ bone marrow are presented from KSL of C/EBPαfl/fl or MxCre+ C/EBPαfl/fl–transplanted mice (right). The numbers represent mean ± SD from 4 similarly transplanted mice. One of 3 experiments giving similar results is presented.

In vivo assay for DC generation from C/EBPα-targeted progenitors. (A) FACS analysis of CD45.2+ donor–derived cells, from 5000 transplanted KSL of C/EBPαfl/fl or Mx1Cre+ C/EBPαfl/fl–transplanted mice, for CDP (Lin, Flt3+, FcγRII/III, cKit+, CD115+, CD11c) from bone marrow, or cDC (CD19, CD3, NK, CD11c+, MHCII+) and pDC (CD19, CD3, NK, CD11c+, B220+) from the spleen show contribution of donor cells to each population. The data shown are representative dot plots from 3 mice per group for 3 independent experiments with similar results. (B) Absolute numbers of total CD45.2+ CD19, CD3, NK CD11c+ DCs were calculated from total splenocytes (left). B220+ subset of cells was determined to be pDC (black), and B220 MHCII+ cells were cDC (white) and plotted accordingly. CMPs were sorted from either MxCre C/EBPαfl/fl or MxCre+ C/EBPαfl/fl mice and analyzed for expression of CX3CR1 by RT-PCR. The percentage of CDP from total CD45.2+ bone marrow are presented from KSL of C/EBPαfl/fl or MxCre+ C/EBPαfl/fl–transplanted mice (right). The numbers represent mean ± SD from 4 similarly transplanted mice. One of 3 experiments giving similar results is presented.

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