Fig. 3.

Detection of cccDNA in tupaia liver biopsy samples. Tupaias were biopsied 4 days before hepatitis B virus inoculation and on days 9, 15, 21, and 42 p.i., and protein-free DNA was extracted from the biopsy samples. (A) The protein-free DNA was subjected to rolling circle amplification (RCA), and the reaction product was linearized by digestion with the restriction enzyme SpeI followed by Southern blotting with the digoxin-labeled HBV DNA-specific probe. (B) cccDNA in protein-free DNA extracted from liver biopsy samples was quantified with quantitative PCR. cccDNA levels in the liver biopsy are displayed as the copy number per microgram of DNA. The figures represent the blotting signal and cccDNA quantification of longitudinal liver tissue from a single HBV-inoculated tupaia. All values are shown as mean ± SD from 3 independent experiments run in duplicate.