FIG 11.

Genome replication, protein expression, and HBsAg and HBeAg secretion following transfection with site-directed mutants of SphI dimer constructs in Huh7 cells. Cells were transfected with the SphI dimer of the parental WT construct, the del3031–3141 or del3033–3131 deletion mutant, or their L-minus (T2936A), S-minus (T156C), L/S-minus (T2936A/T216A), or X-minus (C1632T) mutant. Cells and culture supernatant were harvested 5 days later. (A) Southern blot analysis of intracellular replicative DNA. (B) Western blot analysis of intracellular L, S, HBx, and core proteins, using GAPDH as a loading control. gp19 is a truncated S protein through translation initiation from codon 75 (M75). (C) Western blot analysis of secreted L and S proteins following PEG precipitation. (D) ELISA of secreted HBsAg. (E) ELISA of secreted HBeAg. Data were averaged from three transfection experiments, with the value for the WT construct set as 100%.