FIGURE 2.

HCV protein expression increases PEPCK gene transcription in a CRE-dependent manner. A, luciferase activity of PEPCK-LUC was measured in Huh7, Huh.8, and Ava.1 cells as outlined under “Experimental Procedures.” The luciferase activity was corrected for transfection efficiency. The values are presented as the means ± S.E. *, p < 0.05 (n = 3). B and C, PEPCK RNA levels were measured by qPCR in uninfected control cells and cells infected with either Ad-GFP or Ad-NS5A in both Huh7 cells and isolated primary rat hepatocytes, respectively. The values are presented as the means ± S.E. *, p < 0.05 (n = 3). D, equal amounts of nuclear extracts were collected from HepG2 cells transfected with either Ad-GFP or Ad-NS5A, and subjected to Western blot analysis with NS5A and TATA binding protein (TBP). Representative blots are presented. E, PEPCK mRNA levels were measured by qPCR in cells infected with either Ad-GFP or Ad-NS5A, and the data were normalized to ubiquitin C. *, p < 0.001 (n = 4). F, glucose production was measured in 2-h medium samples of HepG2 cells infected with Ad-GFP or Ad-NS5A (n = 4). The values are presented as the means ± S.E. p = 0.054. G, overexpression of NS5A increased CREB phosphorylation in HepG2 cell line. Equal amounts of nuclear extracts were collected from HepG2 cells infected with Ad-GFP and Ad-NS5A and subjected to Western blot analysis. Representative blots are presented.