Translation efficiencies of identified HCV 5′UTR quasispecies in hepatocytes in primary culture.
Hepatocytes from 3 different donors were used, respectively represented by ■, ▨, and ▪. Approximately 2 × 105 cells were seeded in 24-well plates coated with collagen. After a 3-day culture, they were infected with vTF7-3 recombinant vaccinia virus at 5 PFU/cell for 1 hour at 37°C and then transfected in triplicate as described in “Materials and methods” with respective bicistronic constructs, containing each of the identified HCV quasispecies. At 18 hours after transfection cells were lysed and assayed for luciferases activities. For each quasispecies (QD, QL1, QL2, QP1, QP2, and QP3) IRES relative activities were given as RLuc/FLuc ratios normalized to the QD (originated from DCs) RLuc/FLuc ratio. The data bars and error bars represent the means and SDs of 3 independent triplicate transfections.
