Type I IFN signaling and TLR engagement in early endosomes are unaffected in the absence of DOCK2. (A) Real-time PCR analysis of Irf7 and Ifna4 expression in BM-derived pDCs stimulated with 3 µM CpG-A for the indicated times. Data are expressed as the mean ± SD of triplicate reactions after normalization to expression of the gene encoding hypoxanthine guanine phosphoribosyl transferase 1 (Hprt1) and are representative of two independent experiments. **, P < 0.001; *, P < 0.01. (B and C) BM-derived WT and Dock2^−/− pDCs were stimulated with 3 µM CpG-A (B) or 500 U/ml IFN-α (C) for the indicated times and analyzed for phosphorylation of STAT-1. Data are representative of three (B) and two (C) independent experiments. (D) Real-time PCR analysis of Irf7 and Cxcl10 expression in BM-derived pDCs stimulated with 500 U/ml IFN-α for the indicated times. Data are expressed as the mean ± SD of triplicate reactions after normalization to Hprt1 expression and are representative of two independent experiments. (E–G) BM-derived pDCs retrovirally transduced with TLR9–YFP were stimulated with 3 µM CpG-A for the indicated times. After fixation, cells were stained with ER-Tracker (E), anti–TfR antibody (F), or anti–LAMP-1 antibody (G). The area of TLR9-YFP or CpG-A–Cy5 merged with ER-Tracker (E), TfR (F), or LAMP-1 (G) was calculated in each cell. Bars, 5 µm. (E) Data are expressed as the percentage of colocalization (mean ± SD; n = 10) and are representative of two independent experiments. (F and G) Data were pooled from three independent experiments and are expressed as the percentage of colocalization (mean ± SD) for 30 cells analyzed per group. **, P < 0.001.