Transfection
106 BMDCs from PKR+/+ or PKR-/- mice were primed with ultra-pure LPS for 2 h, and then transfected with 5 μg of Poly I:C by lipofectamine 2000 for 6 h. In another set of experiment, ultra-pure LPS primed BMDCs (106 cells) from PKR+/+ or PKR-/- mice were transfected with 5 μg of Poly I:C or E. coli RNA (Ambion) by electroporation using Nucleofector (Program V-01) and cell line Nucleofector Kit V (Amaxa AG). 3 h later, cell extracts and precipitated supernatants were obtained and analysed by Western blot analysis.