Fig. 5

A moderate effect of HBc core dephosphorylation on genome-containing virion secretion. A Upper panel: Three putative phosphatase-deficient RNase H domain mutants secreted significantly less amount of virion-associated viral DNA than wild type HBV, while HBsAg/virions and naked capsids were similar between wild type and mutants. Middle panel: Extracellular viral and subviral particles were collected from the media of HuH-7 cells on day 5 post-transfection with WT-HBV and mutants. PEG precipitated particles were resolved by native agarose gel, followed by Western blot analyses using anti-core and anti-HBs antibodies. Lower panel: Intracellular capsid particles were analyzed for viral DNA and core by Southern and Western blot analyses. B A bar graph comparison of virion-associated viral DNAs between wild type and three RNase H domain mutants. Signal intensities of HBV DNA associated with mature virions in the top panel of A) were normalized to HBV DNAs associated with intracellular capsids at the bottom panel of A). Signal intensities were quantified by densitometry and an Image J software, *** p < 0.001, ** p < 0.01. p value refers to the Student’s t-test statistical analysis