Inhibition of Interferon Induction and Action by the Nairovirus Nairobi Sheep Disease Virus/Ganjam Virus
Figure 5
Ganjam L protein inhibits transcription from IFNβ-promoter in response to NDV infection.
(a) Vero cells were transfected with 350 ng pIFNβ-luc, 200 ng pJATLacZ and 1.2 µg of a plasmid driving the expression of the indicated viral protein, with the exception of pcDNA6-GV-L1-169 and pcDNA6-GV-N where 700 ng and 300 ng respectively were used in transfection experiments. All transfections were made up to the same amount of DNA using empty vector. After 40 hours of transfection cells were infected with NDV (MOI = 1). After a further 5 hours the cells were (a) lysed with SDS-PAGE sample buffer and the expression of viral proteins detected by Western blot using a monoclonal antibody to the V5 epitope or to the His epitope as required, or (b) lysed with NP40 lysis buffer as described in Methods and the luciferase and β-galactosidase activities determined. Results from two or three separate experiments were combined by setting the RLUs induced by NDV in cells transfected with empty vector to 100%. Error bars show standard errors of normalised data.