Figure 4. Phenotypic analysis of the splenic CD11b⁺ DC subsets.

Unless indicated otherwise, Cx3cr1-GFP reporter mice were used to separate the Esam^hi GFP^lo and Esam^lo GFP^hi subsets of CD11c^hi CD11b⁺ CD8⁻ splenic DCs.

(A) Expression of the indicated surface markers in the gated Esam^hi GFP^lo and Esam^lo GFP^hi subsets. Dashed lines indicate positive staining threshold.

(B) Expression of LTβR on the indicated DC subsets from wild-type mice.

(C) Microphotographs of the sorted DC subsets stained by Giemsa stain on cytospin preparations. The CD11c⁻ CD11b⁺ F4/80⁺ Cx3cr1-GFP⁺ macrophages (MΦ) are shown for comparison (magnification, 400x).

(D) Histograms of BrdU staining in the gated DC subsets after 2 days of in vivo BrdU pulse. The percentages of BrdU⁺ cells are indicated; representative of 4 animals.

(E) Cell cycle profiles of DC subsets after 2 hr of in vivo BrdU pulse. Shown are gated DC subsets stained for DNA content (DAPI) and BrdU incorporation; the fractions of cells in S and G2/M phases are indicated (representative of 2 animals).