Figure 5: Mouse splenic DCs produce type I IFN and cytokines in response to PV-RNAs in a TLR3-dependent manner.

(a) Splenic CD11c⁺ DCs (1 × 10⁶ per ml) isolated from TLR3^−/− or WT mice were stimulated with 20 μg ml⁻¹ poly(I:C) or PV-derived RNAs in FCS-containing medium. Twenty-four hours after stimulation, culture supernatants were collected, and IFN-α/β in the supernatants was quantified using ELISA. TNF-α and IL-6 levels were measured using CBA. Representative data from three independent experiments are shown (mean±s.d.). (b) Poly(I:C) and PV5 (20 μg ml⁻¹) were pre-treated with RNaseIII for 30 min at 37 °C or left untreated before adding to CD11⁺ splenic DCs (1 × 10⁶ per ml) isolated from WT mice. Twenty-four hours after stimulation, TNF-α and IL-6 levels were measured in culture supernatants using CBA (left panels). RNaseIII-treated poly(I:C) and PV5 were electrophoresed on a 2% agarose gel (right panels).