FIG. 2.

Effects of core substitutions on protein expression and DNA synthesis. LMH cells were cotransfected with pCMV-DHBV/C⁻, which expresses a core-defective DHBV genome, together with the indicated core (WT or mutant) expression plasmid. (A) Replicative viral DNA intermediates were isolated (with nuclease digestion [see Materials and Methods for details]) from the cytoplasmic NCs and analyzed by Southern hybridization using a radiolabeled DHBV DNA probe. (B) Core protein expression was analyzed by SDS-PAGE and Western blotting with a core-specific antibody. RC, relaxed circular DNA; SS, full-length single-stranded DNA. The arrowhead indicates the minus-strand DNA (ca. 2.0 kb) reverse transcribed from an internally deleted pgRNA. See the text for details. The marker DNA sizes (kb) are indicated.