HBV core protein allosteric modulators differentially alter cccDNA biosynthesis from de novo infection and intracellular amplification pathways
HBV capsids purified from HepAD38 cells were mock-treated or treated with the indicated concentrations of GLS4, Bay 41–4109 or ENAN-34017 in reactions containing 150 mM NaCl, 50 mM Tris-HCl, pH8.0, 10 mM MgCl2, 1 mM DTT and 0.1% NP-40 at 37°C for 16 h (A) or harvested at the indicated time of incubation (B). Viral DNA were extracted without or with prior DNase I digestion at 37°C for 30 min and detected by Southern blot hybridization with a full-length riboprobe that hybridizes to negative strand of HBV DNA. Partial DS: partially double-starnded DNA. Partial DS: partially double stranded DNA.