FIG. 5.

Increase in mutation-induced core expression is independent of the amount of transfected plasmid DNA. HuH-7 cells were transfected with 20 μg of the control plasmid pGEM7 (lane 1) or 1, 3, 10, and 20 μg of wild-type adwR9 (WTadw; lanes 2 to 5) or MT5/6R9 construct (lanes 6 to 9) together with 19, 17, 10, or 0 μg of pGEM7, respectively (total amount of transfected DNA was 20 μg in all experiments). Core expression was analyzed by SDS-PAGE (15% gel) and immunoblotting of HuH-7 cell lysates with an anticore antibody (A). Quantitation (optical density [O.D.]) of the core protein by using the ImageQuant program is shown below (top row). Transfection efficiencies of wild-type and mutant constructs were similar, as indicated by similar amounts of secreted HBsAg (bottom row). S/N, signal/noise. To demonstrate similar protein loading, the blot was stripped and reprobed with an antiactin antibody (B). Positions of molecular weight markers (in kilodaltons) are indicated on the left.