(a) Structure of BMS-790052
(b) Effect of BMS-790052 on stable replication of a HCV replicon derived from genotype 1b. Huh7 cells harboring replicating subgenomic replicons derived from genotype 1b were treated with 0, 0.1, 1, 10, 100, or 1000 pM of BMS-790052 for 3 days. A total cell lysate from cells was prepared and an equal amount of each cell lysate was separated by SDS-PAGE. Levels of NS5A and β-actin protein expression were examined by western blot analysis using monoclonal anti-NS5A and anti-β-actin antibodies.
(c) Huh7.5 cells were infected with a vaccinia virus expressing a T7 RNA polymerase and then transfected with either Bart79I or Bartman (Con1) plasmid construct containing the NS viral proteins downstream of a T7 promoter. Transfected cells were incubated with 0.1 % of DMSO or 0.01, 0.1, 1, 10, or 100 nM of BMS-790052 for 9 hours. A total cell lysate was prepared and an equal amount of each cell lysate was separated by SDS-PAGE. Levels of NS5A protein expression were examined by western blot analysis using a monoclonal anti-NS5A antibody.
(d) All procedures were performed as in Fig. 1(c) except that levels of NS3 protein expression were examined by western blot analysis using a monoclonal anti-NS3 antibody.