FIG. 2.

Cleavage of K1271 and inhibition of K1271-mediated transactivation by NS3-4A. (A) HEK293 cells were transfected with 100 ng of pRLTK control plasmid, 100 ng of IFN-β-pGL3 reporter plasmid, and 200 ng of K1271 (A) or K1271(A508) (B) Myc-tagged expression plasmid, together with increasing amounts (250 ng, 500 ng, and 1,000 ng) of protease-active NS3-4A or inactive NS3-4A(A139) Flag-tagged expression plasmid as indicated. Luciferase activity was analyzed at 24 h posttransfection. (C and D) The soluble lysates and insoluble fractions were prepared from HEK293 cells used for panels A and B, equilibrated to the same volumes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis loading buffer, and analyzed by immunoblotting with anti-Flag antibody M2 and anti-Myc antibody 9E10.