Fig. 2

Phyllanthus urinaria koreanis extract inhibits HBV DNA synthesis and the secretion of HBsAg and HBcAg. a Southern blot analysis to determine HBV DNA synthesis after treatment with LMV or Phyllanthus extract. At 1 day post-transfection, HepG2 cells were treated with 50 or 100 μg/mL of Phyllanthus extract or LMV (4 μg/mL) for 48 h. HBV DNA was then extracted from isolated core particles, separated, transferred to nylon membranes, hybridized with a random-primed ³²P-labeled HBV-specific probe, and subjected to autoradiography. Single-stranded and double-stranded linear DNA, and partially double-stranded relaxed circular DNA, are marked as SS, DL, and RC, respectively. The graph shows the level of DL DNA relative to that of HBV DNA in the absence of treatment, as measured by the Fujifilm Image Gauge V4.0 program. b Phyllanthus extract inhibits the secretion of HBsAg and HBcAg from HBV WT-transfected HepG2 cells. c RNase protection analysis of HBV cytoplasmic and encapsidated pgRNA from Phyllanthus extract-treated HepG2 cells. The upper panel shows encapsidated pgRNA from isolated core particles, and the lower panel depicts cytoplasmic pgRNA. d Western blot analysis to examine the expression of C protein and core particles in Phyllanthus extract-treated HepG2 cells. Cell lysates were separated by SDS-PAGE and C protein (upper panel) was detected by Western blotting with a rabbit anti-HBc antibody. Isolated core particles were separated on a native agarose gel and analyzed by Western blotting with a rabbit anti-HBc antibody (lower panel). The relative levels of pgRNA, C protein, and core particles were measured using the Fujifilm Image Gauge V4.0 program. Statistical significance was evaluated using Student’s t-test. *p < 0.05 and **p < 0.005, relative to untreated HBV WT