Fig. 2.

CX₃CR1⁺ CD8α⁺ DC lack hallmarks of classical CD8α⁺ DC. (A) Mixed leukocyte reaction with indicated numbers of sorted splenic DC subsets isolated from Cx3cr1^gfp/+ C57BL/6 mice and BALB/c CD4⁺ T cells (10⁵). Data are representative of two experiments. (B) Flow cytometric analysis of splenic DC subsets in 3- and 8-wk-old Cx3cr1^gfp/+ C57BL/6 mice. Bars represent the percentages of CX₃CR1⁻ CD8α⁺ and CX₃CR1⁺ CD8α⁺ subsets out of total cDC (CD11c^hi cells). Data are representative of two experiments. (C) Analysis of IL-12p70 (Left) and IL-12p40 (Right) secretion by sorted splenic DC subsets in response to in vitro exposure to CpG. Data are representative of two experiments.(D) Selective deletion of CX₃CR1⁻ CD8α⁺ splenic DC by CytC injection. Bars represent the percentages of CX₃CR1⁻ CD8α⁺ and CX₃CR1⁺ CD8α⁺ subsets out of total DC (CD11c^hi cells). Data are representative of two experiments. (E) Flow cytometric analysis of splenic DC subsets of Cx3cr1^gfp/+ C57BL/6 mice bearing a WT tumor or a tumor secreting FLt3L. Bars represent the percentages of CX₃CR1⁻ CD8α⁺ and CX₃CR1⁺ CD8α⁺ subsets out of total cDC (CD11c^hi cells) (n = 3). Data are representative of two experiments. (F) Flow cytometric analysis of splenic DC from Batf3^+/+Cx3cr1^gfp/gfp, Batf3^+/−Cx3cr1^gfp/gfp, and Batf3^−/−Cx3cr1^gfp/gfp mice. cDC were gated as CD11c^hi B220⁻ cells.