Figure 3.

Viral markers including cccDNA are reduced on peg-IFNα treatment and rebound after treatment cessation. (A) Experimental setup. HBV-infected USG mice were treated with peg-IFNα as indicated in two independent experiments. (B) Blood was drawn every other week and viral titres were determined by qPCR. The line graph shows mice from experiment 2. Lines depict the median and error bars the range. (C, D) Mice were sacrificed at the indicated time points, and liver DNA extracts (Epicentre-based extraction without proteinase K) were subjected to SB (C). DNA amounts were normalised to human mitochondrial DNA and digested with PSD before loading. The bar graph below shows the densitometry analysis of the cccDNA band. The blot shows mice from experiment 2. (D) cccDNA levels were determined by qPCR in Epicentre-based DNA extracts without proteinase K after PSD digestion in all mice (same extracts as in B). cccDNA copies were normalised to the human mitochondrial gene ND2. Each dot represents a single mouse; horizontal lines depict the median. cccDNA, covalently closed circular DNA; MyrB, myrcludex-B; NT, not treated; IFNα, interferon-α; MyrB, myrcludex-B; NT, not treated; peg-IFNα, pegylated interferon-α; pf-rcDNA, protein-free relaxed circular DNA; qPCR, quantitative PCR; SB, Southern blot; USG, uPA/SCID/beige/IL2RG^–/–.