Hepg2

HBMECs (human brain microvascular endothelial cells) and HUVECs (human umbilical vein endothelial cells) were purchased from DS Pharma Biomedical (Osaka, Japan). HBMECs and HUVECs were cultured in collagen-coated plates in HuMedia EG-2 (Kurabo, Osaka, Japan). HCF (Human cardiac fibroblasts) were purchased from ScienCell Research Laboratories (Carlsbad, CA). HCF were cultured in RPMI 1640 medium with L-glutamine and 10% FBS. hNHeps™ (Human normal hepatocytes) cells were purchased from Lonza (Lonza, Walkersville, MD). hNHeps™ cells were maintained in hepatocyte culture medium (Lonza). A549 (human lung adenocarcinoma) and HeLa (human epithelial cervical cancer) cells were purchased from Sigma-Aldrich (St. Louis, MO). HepG2 (human hepatocellular carcinoma) cells were provided from Cosmobio (Tokyo, Japan). Caco-2 (human colonic carcinoma) cells were purchased from American Type Culture Collection (Rockville, MD). A549, HeLa, HepG2, and Caco-2 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Wako Pure Chemical Industries, Osaka, Japan) containing 50 units/ml penicillin and 50 μg/ml streptomycin (Sigma-Aldrich, St. Louis, MO), and supplemented with 10% FBS (Wako Pure Chemical Industries). These cells were cultured at 37°C in a humidified atmosphere of 5% CO₂.

Human hepatoma cell lines HepG2 and Huh-7 cells were obtained from American Type Culture Collection (Manassas, VA, USA) and the Health Science Research Resources Bank (Osaka, Japan), respectively. HepG2 and Huh-7 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) with a low glucose concentration (1 g/L) (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented by 10% fetal calf serum (FCS) at 37 °C in a humidified atmosphere containing 5% CO₂. Human embryonic kidney (HEK) 293T cells were maintained in DMEM with a high glucose concentration (4.5 g/L) (FUJIFILM Wako Pure Chemical Corporation) supplemented by 10% FCS at 37 °C in a humidified atmosphere containing 5% CO₂. HepG2-NTCP cells were generated by inoculating HepG2 cells with a lentiviral vector which contains NTCP and neomycin-resistance genes. The transduced cells were selected by 600 μg/ml G418 (Tokyo chemical industry, Tokyo, Japan) and prepared for HBV inoculation experiments. Human primary hepatocytes, PXB-cells (PhoenixBio, Hiroshima, Japan), were maintained in the culture medium for PXB-cells (PhoenixBio) according to the manufacturer’s instructions. Huh-7/HBc-FLAG cells were transduced with a lentiviral vector (pLVSIN-EF1α-Hyg, TaKaRa) containing TRIM26ΔR and selected by 250 μg/ml hygromycin B to establish the cells stably express TRIM26ΔR (Fig. 7B).