Differentiation and maturation of DCs are normal in Tyk2^−/− mice. (A) Splenocytes from wild-type (WT) mice and Tyk2^−/− mice were stained with anti-CD11c FITC, anti-CD11b PE, anti-B220 PerCP, anti-CD3 PerCP, and anti-CD8 allophycocyanin or with anti-CD11c FITC, DX-5 PE, anti-CD11b PE, TER-119 PE, CD19 PE, anti-CD3 PerCP, and anti-B220 allophycocyanin, and the stained cells were analyzed by FACS. The frequencies of CD11b⁺ DCs (CD11c⁺ CD11b⁺ B220⁻ CD3⁻ CD8⁻ cells), CD8⁺ DCs (CD11c⁺ CD8⁺ CD11b⁻ B220⁻ CD3⁻ cells), and plasmacytoid DCs (PDCs) (CD11c⁺ B220⁺ CD3⁻ CD11b⁻ CD19⁻ cells) are shown. Data are means (± SD) for 5 mice in each group. (B) Isolated CD11c⁺ DCs from WT splenocytes or Tyk2^−/− splenocytes were cultured with CpG ODN (10 μg/mL) for 3 days and analyzed for the expression of I-A^d and CD80 by FACS. As controls, freshly isolated CD11c⁺ DCs from WT splenocytes or Tyk2^−/− splenocytes were analyzed for the expression of I-A^d and CD80. Shown are representative histograms from 4 independent experiments. Dashed lines indicate the staining with isotype-matched control antibodies. (C) Isolated CD11c⁺ DCs from WT splenocytes or Tyk2^−/− splenocytes were subjected to intracellular staining with anti-TLR9 antibody (bold lines) or control antibody (dashed lines). Shown are representative histograms from 3 independent experiments.