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. 2021 Jan 28;71(2):372–381. doi: 10.1136/gutjnl-2020-322571

Figure 1.

Figure 1

Anti-HBV siRNA treatment potently reduces all HBV markers except cccDNA. (A) Experimental setup. HBV-infected USG mice were treated as indicated and blood was drawn every other week. (B–D) Viral titres were determined by qPCR (B), and serum HBsAg (C) and HBeAg (D) were determined by ELISA (Abbot Architect). Every line represents one mouse. (E, F) Mice were sacrificed at the indicated time points, and cccDNA levels were determined by qPCR in Epicentre-based DNA extracts without proteinase K after PSD digestion (E). cccDNA copies were normalised to the human mitochondrial gene ND2. Each dot represents a single mouse; horizontal lines depict the median. (F) Liver DNA extracts (Epicentre-based extraction without proteinase K) from D were subjected to Southern blot. 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 depicts one representative experiment. cccDNA, covalently closed circular DNA; MyrB, myrcludex-B; NT, not treated; pf-rcDNA, protein-free relaxed circular DNA; qPCR, quantitative PCR; SB, Southern blot; siRNA, small interferring RNA; USG, uPA/SCID/beige/IL2RG–/–. †=this mouse died prematurely at the 2-week blood drawl.