FIGURE 7.

HCV replication assay. Huh7 (bar 1) and FCA4 lysates containing RNP complexes hybridized with oligonucleotides and followed by the avidin-agarose affinity purification step were incubated with [³²P]CTP for 90 min during an RNA synthesis reaction, as described in Materials and Methods (bar 4). The incorporation of ³²P radioactivity in the FCA4 lysates directly subjected to RNA synthesis (bar 2) or annealed to the neomycin oligonucleotide followed by RNA synthesis (bar 3) is presented. The radiolabeled lysates (1 μL) were spotted on DE81, ion exchange filters (Whatman). The filters were dried and washed extensively with 100 mM sodium phosphate buffer (pH 8.0). The ³²P-labeled RNA products bound to the filters were measured by scintillation counting.