FIG. 10.

(A) NS3-4A and A20 inhibit K1271-mediated activation of endogenous ISG56 gene expression. HEK293 cells were cotransfected with 1 μg of pcDNA3, Myc-ΔRIG-I, Myc-KIAA1271, Myc-KIAA1271(C508A), or Myc-TRIF and 2 μg of IRF-3ΔN, Flag-A20, or Flag-NS3-4A expression construct as indicated. Whole-cell extracts (50 μg) were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and analyzed by immunoblotting using antibodies to ISG56, Myc, Flag, IRF-3, and actin. (B) K1271 inhibits VSV replication. HEK293 cell lines expressing K1271 or IKKɛ were infected with VSV (1 multiplicity of infection), and viral protein expression was measured at different times after infection by immunoblotting. Expression of the Flag-tagged transgene and the endogenous ISG56 was also monitored.