Formation of tubular endolysosomal structures was impaired in pDCs and macrophages derived from Ly49Q knockout mice. (A) Bone marrow–derived pDCs were prepared by culturing bone marrow cells with Flt3L, and they were then incubated with rhodamine-conjugated CpG (0.3 μM) for 60 minutes. The cells were fixed and then stained with an anti–LAMP-1 antibody. Confocal images of representative cells are shown. (B) The number of cells showing the directionally extending CpG-including tubular endolysosomal structures was counted, and the proportion of total cells was calculated. (C) Peritoneal exudate macrophages were prepared from Ly49Q knockout and control littermate mice and incubated with rhodamine-conjugated CpG (0.3 μM) for 24 hours. The intracellular distribution of CpG was examined by confocal microscopy. (D) The peritoneal exudate macrophages were treated with rhodamine-conjugated CpG (0.3 μM) for 24 hours. The cells were fixed and then stained with anti–detyrosinated tubulin (Glu-tubulin) antibody. Arrowheads indicate MTOC. (E) Cytokine production by peritoneal exudate macrophages was compared between Ly49Q-deficient and control littermate mice. Macrophages were stimulated with CpG, and the amount of cytokine in the culture supernatant was estimated by cytokine bead array 24 hours later.