Figure 2: Cytokine expression profiles of pDCs treated with low or high doses of CpG or GpC.

(a) Il6, Il10, Il23a, Ifna1, Ifnb1, Ifng and Tgfb1 transcripts were evaluated in pDCs treated with 1 or 10 μg ml⁻¹ of CpG, nc-ODN or GpC for 6 h. IL-6, IL-10, IL-23 p19, IFN-α, IFN-β, IFN-γ and TGF-β mRNAs were quantified by real-time PCR using Gapdh normalization. Data (means±s.d. from four experiments) are presented as transcriptional expression of the different cytokines relative to basal cytokine transcript amount in unstimulated cells. *P<0.005; Student’s t-test. (b) pDCs were assayed for cytokine release after CpG, nc-ODN or GpC exposure. Data are means±s.d. from four experiments. (c) Intracellular TGF-β staining profiles of B220⁺ pDCs treated with 1 or 10 μg ml⁻¹ of CpG for 24 h. Shown in upper right quadrants are percentages of double-positive cells. One experiment is shown representative of six, all of which are visualized by scatter plot in Supplementary Fig. S6. (d) Production of TGF-β by pDCs untreated or treated with 1 (pDC/L-CpG) or 10 (pDC/H-CpG) μg ml⁻¹ of ODN. Supernatants were harvested at 72 h after stimulation and assayed for active TGF-β. *P<0.01; Student’s t-test.