Fig 2.

GBVA10-9 does not affect HCV attachment or viral RNA replication. (A) HCVcc was mixed with GBVA10-9 (2 μM), C5A (8 μM), scrambled peptide (2 μM), or the HCV attachment inhibitor heparin (200 μg/ml) and then added to precooled Huh7.5.1 cells and incubated for 2 h at 4°C. The cells were then thoroughly washed in PBS and subjected to RNA isolation. Cell surface-bound HCV was quantified by qRT-PCR. **, P < 0.005. (B) After virus attachments and washes at 4°C as described above, a set of parallel cells were moved back to 37°C and incubated for 2 additional days. The intracellular HCV RNA titer was measured. (C) GBVA10-9 (2 μM), scrambled peptide (2 μM), or IFN-α2b (100 U/ml) was added to 2⁻3⁺ cells for 12 h before the medium was changed. At 72 h postinfection, intracellular HCV RNA was quantified by qRT-PCR. (D) Huh7.5.1 cells were infected with HCVcc (multiplicity of infection = 0.01) for 12 h and then treated with GBVA10-9 (2 μM), scrambled peptide (2 μM), DMSO (0.5%), PBS, or IFN-α2b (100 U/ml) for an additional 3 days, during which fresh peptides or IFN-α2b was added every 24 h. At 72 h postinfection, immunoblotting was performed with an anti-HCV core antibody.