FIG. 5.

Protein expression in the presence of a trans-lactam. Compound 1b was added, at the concentrations indicated, to BT7H cells at different stages during the labeling process. The 21-h samples were continuously exposed to the compound. The 3-h sample was exposed to the compound from 18 to 21 h postinfection, during both the methionine starvation and the labeling phase. The 2-h sample was exposed to the compound from 19 to 21 h postinfection, during the metabolic labeling phase only. SDS-PAGE was carried out on the labeled samples. Western blot analysis with an in-house anti-NS3 polyclonal antibody was performed on parallel samples to assess the total amount of NS3 in the system. (A) Autoradiograph showing metabolically labeled proteins. MW, molecular weight marker (in thousands). P, precursor proteins. (B) Western blot analysis with the anti-NS3 antibody.