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. 2007 Apr 4;81(12):6164–6174. doi: 10.1128/JVI.02721-06

FIG. 2.

FIG. 2.

Formation of HBV and DHBV CCC DNA in stably transfected hepatoma cell lines. HBV (A) and DHBV (B) DNAs were extracted from the cytoplasmic nucleocapsid (core) with protease digestion or the Hirt supernatant without protease digestion (PF), respectively, from HepAD38 (HBV) and Dstet5 (DHBV) cells cultured in the absence of Tet and analyzed by Southern blotting. The nature of CCC DNA in the HBV PF sample (panel A, lanes 5 and 6) was further confirmed by heating to 95°C, which denatured RC/NC and DSL DNA to SS without denaturing CCC DNA, or at 95°C, followed by EcoRI digestion, which linearized CCC DNA. Note the predominance of CCC DNA and RC/NC DNA in the DHBV and HBV PF DNAs, respectively. El, EcoRI. (C) The kinetics of accumulation of CCC DNA and PF-RC DNA in HepAD38 (HBV) and Dstet5 (DHBV) cells. The PF DNA was extracted after induction for 7, 10, 14, and 17 days for HBV (lanes 1 to 4) and 4, 5, 6, and 7 days for DHBV (lanes 5 to 8) and analyzed by Southern blotting.