FIG. 5.

Effect of L-AGL mutations on HDV infectivity. SL-HDV particles bearing mutations in L-AGL were produced in Huh-7 cells. Mutations included a deletion of residues 119 to 123 (Δ119-123) in the S domain of L-HBsAg, or single amino acid substitutions as indicated. (A) Particles from 0.5 ml of cell culture supernatant were analyzed for S- and L-HBsAg proteins by immunoblotting. The glycosylated (gp) and nonglycosylated (p) forms of S- and L-HBsAg are indicated. Genomic HDV RNA from 140 μl of supernatant was analyzed by Northern blotting. (B) After normalization of the different preparations of HDV particles for their viral RNA content, they were assayed for infectivity in cultures of HepaRG cells. Infectivity was evaluated by measuring the accumulation of HDV RNA in cells harvested at day 7 postinoculation. Quantification of HDV RNA signals by phosphorimager is indicated as percentages of the value for wt SL-HDV. The experiment was repeated with consistent results.