FIG 2.

Antiviral mechanism of BA derivatives against HBV. AML12HBV10 cells were cultured in the presence of tetracycline (tet +) or in the absence of tetracycline and mock treated (tet −) or treated with the indicated concentrations of the compound BA-26019 (1 and 3 μM), BA-38017 (1 and 3 μM), ETV (1 μM), DVR-23 (2 μM), or Bay 41-4109 (2 μM) for 2 days. (A) Intracellular viral RNA was measured by Northern blotting hybridization. 28S and 18S rRNA were used as loading controls. (B) HBV core protein (Cp) expression was detected by Western blotting with a rabbit polyclonal antibody (Dako). β-Actin served as the loading control. The results were derived from two separate gels, as indicated. (C) Encapsidated pgRNA was determined by Northern blotting. (D) The total amounts of capsids were determined by a particle gel assay in a 1.0% agarose gel electrophoresis. Capsid-associated HBV DNA was detected by hybridization upon alkaline treatment of nucleocapsids on the membrane following the particle gel assay. (E) HBV DNA replication intermediates were extracted and determined by Southern blotting. A ³²P-labeled full-length minus-strand-specific riboprobe was used for Southern blot analysis. RC, relaxed circular DNA; DSL, double-stranded linear DNA; SS, single-strand, negative-polarity DNA.