Fig. 7.

The E3 dsRNA-binding domain blocks IRF3 and NF-κB activation in response to poly(dA–dT) transcribed RNA. 293T cells were co-transfected with pRL-TK and either (a) ISG56.1 or (b) NF-κB reporter plasmids together with plasmids expressing either E3 or the E3 dsRNA-binding domain or the empty vector control (pcDNA4) overnight. The cells were then transfected with 1 μg total RNA extracted from 293Ts that had been transfected with 12 μg poly(dA–dT) for 24 h. At 24 h post-transfection, cells were lysed for dual luciferase reporter assay. (c) As a positive control 293T cells were co-transfected with the NF-κB reporter plasmid as described for (b) and the following day were transfected with 800 ng poly(I : C) for 24 h prior to harvesting of cells for dual luciferase reporter assay. Error bars indicate the mean±sem (*P<0.05, **P<0.01, ***P<0.001).