Figure 1

HCV replication in human hepatocytes. (A) Intracellular distribution of PTEN: Cells were visualized at day 3 post-transfection with HCV genomic RNA. The uninfected cells (left four panels) show uniform PTEN staining; HCV replication is marked with NS5A antigen (far right panel). Transfection efficiency of HCV genomic RNA was approximately 30%. (B) PTEN protein in HCV-infected cells: 1 × 10⁵ PPH cells were transfected with 1 μg H77 HCV1a genomic RNA, and the cells were collected at day five (U, uninfected; I, infected). Total cell protein was extracted using RIPA buffer; 20 μg of protein was analyzed on 10% precast Protein Gel; numbers underneath each lane indicate the relative values of PTEN. HCV replication is marked with viral core antigen (HCV Core). (C) Restriction of nuclear PTEN protein in HCV-infected cells: 1 × 10⁵ PPH cells were transfected with increasing amounts (0.5, 1.0, 2.0 μg) of input HCV1a genomic RNA, cells were harvested 5 days post-transfection and 10 μg each of nuclear or cytoplasmic protein were analyzed by Western blot. GAPDH and Lamin B1 were loading controls respectively, for the cytoplasmic or nuclear protein.