Fig 6.

vIRF1 or vIRF2 expression leads to increased EMCV viral production following poly(I·C) pretreatment. 293-TLR3 cells were transfected with the control vector or vectors expressing vIRF1, -2, or -3. Twenty-four hours posttransfection, cells were treated with poly(I·C) for 6 h prior to infection with EMCV at an MOI of 0.001 for 24 h before harvesting. (A) Quantitative RT-PCR was performed on RNA to investigate EMCV genetic material, normalized to that of β-actin, represented as the fold increase over vector mock-infected cells. (B) Plaque assays were performed on L929 cells with six different 1:10 serial dilutions of viral supernatant. (C) Lysates harvested at the time of EMCV infection were subjected to SDS-PAGE and immunoblotted with the epitope tags Myc (vIRF1), Xpress (vIRF2), and FLAG (vIRF3) to determine the expression levels of vIRFs posttransfection. Values represent the means plus or minus the standard deviations of the means from triplicate biological replicates. *, P < 0.02 (Student's t test).