B220⁺CD19^-Ly-6C⁺CD11c^Lo cells in bone marrow are functional pDCs. (A, left) Whole bone marrow (wBM) from BALB/c mice was stained for CD45R/B220, Ly-6C, CD19, and CD11c. The B220⁺CD19^- population, gated with an open box, was further resolved and sorted according to CD11c and Ly-6C expression (middle). (Right) Purity of sorted cells. (B) The sorted B220⁺CD19^-Ly-6C⁺CD11c^Lo cells were stimulated with CpG-ODNs for 12 hours, and their morphology was assessed by May-Grünwald-Giemsa staining. (Top) Unstimulated cells; (bottom) stimulated cells. (C) Culture supernatants were analyzed for cytokine levels by ELISA after stimulation with CpG-ODNs (▪) or not (▦). These results are representative of those obtained in 4 independent experiments.