Fig 8.

Role of caspase cleavage in NS5A nuclear localization and HCV RNA replication. (A) Schematic representation of lentivirus vectors carrying mutated NS5A-v1. The closed triangle indicates mutation of the caspase recognition site at amino acid position 154. LTR, long terminal repeat. (B) Confocal analysis of cells transduced with the mutated NS5A-v1 lentiviruses was carried out as described in the legend to Fig. 4. Scale bar, 10 μm. *, nontransduced cells. The yellow arrow shows where the 3D reconstructed image was cropped to display the cell sections. The yellow pictogram indicates the direction of sight. (C) Effect of caspase cleavage of NS5A on HCV replicon replication. (Upper panel) Schematic representation of three subgenomic replicons encoding NS5A-v1 with a mutated caspase cleavage site. An open triangle denotes mutation of the caspase recognition site at amino acid position 154; a closed triangle indicates the presence of the cell culture adaptive mutation S2204I. (Lower panel) Replication efficiency of the indicated subgenomic replicons was analyzed in the presence or absence of the caspase inhibitor z-VAD-fmk at the indicated concentrations. The data represent the mean ± SEM replication efficiencies obtained from 3 independent experiments carried out in triplicate.