Adefovir

[³H] Adefovir (11.9 Ci/mmol) and [¹⁴C] Inulin methoxy (6.2 mCi/g) (Moravek Biochemicals Inc., Brea, CA), Inulin, Adefovir, and urethane (Sigma-Aldrich, St. Louis, MO), 20% mannitol (Baxter Healthcare Corporation, Deerfield, IL), Ultima GOLD and Solvable (Perkin Elmer, Waltham, MA). Radiolabeled Adefovir was used to facilitate detection throughout the course of the study, and because scintillation counting would not be complicated by metabolite presence. The rate of decomposition of radiolabeled compounds guaranteed by supplier is approximately 1%/month for the first six months after purification when stored at -20 °C. Both radiolabeled compounds were therefore used within the 1 month after delivery from supplier.

Chicken hepatoma cells (LMH) were cultured in DMEM/F12 medium (Mediatech) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin. dstet5 is a LMH-derived stable cell line supporting the replication of an envelope protein-deficient DHBV genome in a tetracycline dependent manner (Guo et al., 2003 (link)). HepAD38 is a human hepatoma cell (HepG2)-derived stable cell line supporting HBV genome replication in a tetracycline dependent manner (Guo et al., 2007 (link)). Both cell lines were maintained in DMEM/F12 medium (Mediatech) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 1 μg/ml tetracycline and 200 μg/ml G-418. Chicken alpha-interferon (IFN-α) was produced and titrated as described previously (Guo et al., 2003 (link)). Foscarnet (PFA), Lamivudine (LAM), and Adefovir (ADV) were purchased from Sigma. Entecavir (ETV) was provided by Dr. William S. Mason at Fox Chase Cancer Center, Philadelphia. Bay 41-4109 was provided by Dr. Lai Wei at Hepatology Institute of Peking University, China. AT-61 and DVR-23 are synthesized in house as described previously (Campagna et al., 2013 (link); Xu et al., 2010 (link)).