Figure 2
From: Replication of a hepatitis C virus replicon clone in mouse cells
HCV protein and RNA detection in immortalized mouse hepatocytes. (A) Western Blot detection of HCV NS5A. Cell lysate was harvested from individual cell clones and resolved by SDS-PAGE. Samples include cell lysate from Huh-7 sgCon1 (lane 1); non-transfeted (NT) Huh-7 (lane 2); Huh-7 sgJFH-#2 (lane 3); NT MMHD3 cells (lane 4); MMHD3 sgJFH#1 (lane 5); and MMH1-1sgJFH#4, 5, 6, 7, 9, and 10 (lanes 6–11). (B) Northern Blot detection of sgJFH-1 RNA. Total RNA was isolated from non-transfected (NT) and individual MMHD3 (lanes 2), MMH1-1 (lanes 4–9), and Huh-7 (lanes 11–15) sgJFH-1 replicon clones. Positive-strand (top panel) and negative-strand (middle panel) HCV RNA was detected with 32P-labeled strand-specific riboprobes. Cellular GAPDH was detected with a 32P-labeled cDNA probe (bottom panel). Serial dilutions of in vitro transcribed positive-strand and negative-strand sgJFH-1 RNA (109, 108, 107) are shown in lanes 16–18 and lanes 19–21, respectively.