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. 2024 Jan 5;98(2):e01721-23. doi: 10.1128/jvi.01721-23

Fig 3.

Fig 3

Effects of Tet deletion on liver viral transcript and replication intermediate levels in HBV transgenic mice. Quantitative analysis of (A) HBV 3.5-kb RNA and (B) total HBV RNA (3.5-kb plus 2.1-kb RNA) by NanoString gene expression analysis in HBV transgenic mice. Individual (circles) and mean (line) transcript levels are indicated for male (M), female (F), homozygous Tet2(fl/fl)Tet3(fl/fl) alleles (Tet2 and Tet3), Cre-negative allele [Cre(−)], and Cre-positive allele [Cre(+)] HBV transgenic mice. Statistical differences (adjusted P values; Tukey’s multiple comparison test) in the levels of the transcripts between Cre(−) and Cre(+) HBV transgenic mice are indicated (GraphPad Prism 10.0.3). Transcript levels from adult Tet-deficient HBV transgenic mice (left panel) are compared to the changes in gene expression observed throughout postnatal liver development in wild-type HBV transgenic mice (right panel). Arrows indicate the average levels of transcript observed in HBVTet2(fl/fl)Tet3(fl/fl)Cre(−) (top arrow) and HBVTet2(fl/fl)Tet3(fl/fl)Cre(+) (bottom arrow) mice. (C) RNA (Northern) filter hybridization analyses of liver transcripts isolated from HBV transgenic mice are shown. Noncontiguous lanes from a single analysis are presented. The probes used were HBVayw genomic DNA plus Gapdh cDNA. The Gapdh transcript was used as an internal control for the quantitation of the HBV 3.5-kb RNA. Serum HBeAg levels encoded by the HBV 3.5-kb precore RNA are also indicated. (D) DNA (Southern) filter hybridization analyses of liver replication intermediates isolated from HBV transgenic mice are shown. Noncontiguous lanes from a single analysis are presented. The probe used was HBVayw genomic DNA. The HBV transgene (Tg) was used as an internal control for the quantitation of the HBV replication intermediates. RC, HBV relaxed circular replication intermediates; SS, HBV single-stranded replication intermediates.