Fig. 1.
Production of HBV transgenic mice with liver-specific knockout of Atg5. (A) Schematic illustration for the production of HBV transgenic mice with liver-specific knockout of Atg5 and their control littermates. The HBV transgenic mouse line Tg05 has been described previously (21). The floxed Atg5 mouse line was a generous gift of Noboru Mizushima (5), and the Alb-Cre mouse line was obtained from Jackson Laboratory (13). All three mouse lines had the C57BL/6 genetic background. The final breeding step also produced HBV+/− Atg5f/f control mice without Alb-Cre (not shown). The use of either Alb-Cre HBV+/− Atg5+/f or HBV+/− Atg5f/f control mice generated similar results for HBV. HBV ATG5-WT, HBV transgenic mice with the wild-type ATG5 gene; HBV L-ATG5-KO, HBV mice with liver-specific knockout of the ATG5 gene. (B) Western blot analysis for autophagy in the mouse liver. ATG5 represents the conjugated heterodimer of Atg5-Atg12, as ATG5 was not detectable by itself in mouse livers (16). LC3-I and LC3-II are nonlipidated and lipidated LC3, respectively. Lipidated LC3 has a higher electrophoretic mobility than nonlipidated LC3 (18). The p62 protein was analyzed for measuring the autophagic protein degradation, and α-actin served as the loading control. Two 2-month-old male mice were used in each group for confirmation of the results.