Figure 6
Expression of HCV polyprotein from VV inhibits cellular and viral protein synthesis in the hepatic cell line HepG2. A: Monolayers of HepG2 cells were infected (5 PFU/cell) with either VT7 or VT7-HCV7.9 recombinant viruses, in the presence (+) or absence (-) of the inductor IPTG. Uninfected (U) and infected cells were metabolically labelled with [35S] Met-Cys Promix (100 μCi/mL) from 4 to 24 h.p.i and treated as described under Materials and Methods. Approximately 100 μg of total cell protein extracted from uninfected and infected cells was fractionated by SDS-PAGE followed by autoradiography. (*) represents new additional polypeptides corresponding to the HCV proteins. B: Immunoblot analysis of total cell protein lysates prepared from uninfected and infected cells at 24 h.p.i. The blot was probed with a rabbit polyclonal anti-serum raised against live VV. C: The blot was stripped and probed again with a polyclonal antibody that recognized phospho-eIF-2α-S51 protein.