Figure 1.

Immunostimulatory RNA production can be detected 7 hours after VSV infection and parallels both IFN-β induction and translation of viral proteins in infected cells. HEK 293 cells were infected with VSV (MOI=1). At the indicated time points post infection cells were lysed, RNA isolated and cytoplasmic and nuclear protein fractions were prepared. (A) RNA from infected cells was analyzed by RT-qPCR for the expression of IFN-β. (B) Expression of VSV G, IRF3 and β-actin in the cytoplasm and phosphorylated IRF3 in the nuclear fraction was determined by western blotting. (C) RNA isolated from VSV-infected HEK 293 cells at the indicated time points after infection was retransfected (100 ng/100 µl) into 1205Lu melanoma cells (8 x 10³ cells/well) using RNAiMax Lipofectamin. IP-10 was measured in the supernatant after 24 hours. Data are shown in (A, C) as mean +- SD of triplicates and are representative for n=2 independent experiments.