Ly49Q colocalized with CpG in endosome/lysosome compartments. (A) Colocalization of Ly49Q with CpG in endosomes. Peritoneal exudate macrophages were prepared from Ly49Q Tg mice. The cells were incubated with rhodamine-conjugated CpG (0.3 μM) for 15 minutes, fixed with 4% formalin in PBS, then stained with an anti-FLAG antibody and analyzed by confocal microscopy. The bottom 4 photographs show serial Z-axis–sectioned patterns after 60-minute incubation with rhodamine-conjugated CpG. Squares indicate the region shown in higher magnification. (B) Localization of CpG to late endosomes/lysosomes. RAW264 cells were incubated with rhodamine-conjugated CpG for 60 minutes. To visualize the late endosomes/lysosomes, the cells were incubated with lysotracker for the last 30 minutes. (C) Localization of Ly49Q in the late endosomes/lysosomes. A Ly49Q-DsRed fusion construct was introduced into RAW264 cells. Twenty-four hours after transfection, the cells were cultured with lysotracker at 37°C for 30 minutes, and the intracellular localization of Ly49Q was examined by confocal microscopy. Ly49Q localized to the late endosomes/lysosomes. In the presence of CpG stimulation, Ly49Q-containing compartments showed a tubular structure.