Skip to main content
. 2008 Oct 1;82(23):11927–11938. doi: 10.1128/JVI.00924-08

FIG. 5.

FIG. 5.

Effect of LaR2C-N7 on HCV IRES-mediated translation in vitro. (A and B) Above the panels is a schematic representation of the wt 7-mer peptide (LaR2C-N7) and the mutant 7-mer peptides, along with the 24-mer wt LaR2C peptide. The mutated residue is indicated in italics. One microgram of HCV IRES Luc monocistronic RNA was translated in RRL in the absence or presence of increasing concentrations (15, 30, and 60 μM) of either the wt LaR2C-N7 or the mutant mLaR2C-N7-4 peptide. Respective luciferase activities were measured and plotted against different peptide concentrations. The relative FLuc activities are represented as percentages of the control reaction (expressed as 100%). Results represent an average of three independent experiments. (C) Similarly, 1 μg of capped luciferase RNA was translated in RRL in the absence or presence of increasing concentrations (15, 30, 60 μM) of LaR2C-N7 peptide, and luciferase activities were plotted against different concentrations of the wt LaR2C-N7 peptide. The relative FLuc activities were represented as percentages of the control reaction (expressed as 100%). Results represent an average of three independent experiments. (D) One microgram of capped bicistronic RNA was translated in RRL in the absence or presence of increasing concentrations (15, 30, 60 μM) of LaR2C-N7 peptide, and luciferase activities were plotted against different concentrations of the wt LaR2C-N7 peptide. The relative RLuc and FLuc activities were represented as a percentage of the respective control reactions (expressed as 100%). Results represent an average of three independent experiments. (E) One microgram of either PV-luciferase monocistronic RNA or capped HAV-bicistronic RNA (containing Rluc-HAV-Fluc, in order) translated in the absence (lane 1) and presence of increasing concentrations (15, 30, 60 μM) of the LaR2C-N7 peptide. The translation of the firefly luciferase activities was measured and plotted relative to the peptide concentrations for the respective constructs as indicated. The relative FLuc activities were represented as percentages of the control reaction (expressed as 100%). Results represent an average of three independent experiments. (F) One microgram of HCV IRES Luc monocistronic RNA was translated in RRL in the absence or presence of wt LaR2C-N7 peptide (40 μM). Increasing concentrations (25 ng and 50 ng) of purified wt La protein or bovine serum albumin (50 ng) were added to the reaction mixtures as indicated below the lanes. Respective luciferase activities were measured and plotted in the graph. The relative FLuc activities were represented as percentages of the control reaction (expressed as 100%). Results represent an average of three independent experiments.