FIG 4.
ABI-H0731 disrupts pgRNA encapsidation and HBV DNA replication. Shown are data from comparative analyses of the inhibitory effects of ABI-H0731, other CIs, and ETV at concentrations 10-fold over their VR EC50s on HBV infection markers of induced HepAD38 cells. (A) HBV pgRNA and surface RNAs (sRNAs) were detected from total RNA extracts via Northern blotting, with 18S and 28S rRNAs serving as loading controls. Levels of encapsidated RNA were detected from the micrococcal nuclease (MNase)-treated cell lysate by Northern blotting. (B, top) Disruption of capsid formation was examined in EIA gels. (Middle and bottom) Levels of HBV core protein (middle) and GAPDH (control) (bottom) were evaluated by Western blotting using total cell extracts. (C) Levels of total capsid-associated viral DNA (top) and the extracted HBV DNA replication intermediates (bottom) were assessed by Southern blotting. Band density was quantified using ImageJ software. rcDNA, relaxed circular DNA; ssDNA, single-stranded DNA.