Fig. 4
DI mutants and their influence on translation using the Luc-SG replicon.a Illustration of the HCV 5′ UTR and IRES and location of the inserted mutations (orange). b Sequence alterations of DI mutants compared to WT. The miR-122 seed binding sites are marked by green boxes. Note that all mutants lack wild type miR-122 binding, due to mutations in the seed sequence. c Translation assays in Hep3B cells, using firefly luciferase (FLuc) reporter replicons and a capped Renilla (RLuc) control. miRmut or miRWT were co-electroporated, and luciferase activity was measured after 1, 2 and 4 h. All constructs were replication deficient ΔGDD mutants, to exclude potential effects of early replication. Mutations are described in the “mut” row, and a pictogram of the predicted minimum free energy structure in the “folding” row. Mutants enhancing SLII formation are depicted in green, SLIIalt stabilizing in red. Mean values (±SD), n = 3, duplicates. RLU relative light units. Statistical significance of the difference between WT and DI mutants in the presence of miRmut is indicated. The reference graph used to calculate is given in each subpanel in light gray. *P < 0.05, **P < 0.01, ***P < 0.001
