Hcc36

Male BALB/c nu/nu mice were obtained from the National Laboratory of Animal Breeding and Research Center (Taipei, Taiwan) and housed according to the protocols of the Animal Center, Kaohsiung Medical University, Kaohsiung, Taiwan [37 (link)]. Normal hepatocyte cells (Chang normal liver cells; CNL) and human hepatoma cell lines (HepG2, Hep3B, SK-Hep1, Huh7, PLC/PRF/5, HCC 36 and HA22T) were obtained from American Type Culture Collection (Manassas, VA, USA) and maintained according to the manufacturer's protocol [38 (link)].

Human hepatocellular carcinoma cells HCC36 (American Type Culture Collection, Manassas, VA, United States) were cultured in Dulbecco’s Minimal Essential Medium (Sigma-Aldrich, Burlington, MA, United States) supplemented with 10% heat-inactivated bovine calf serum, 100 units/ml penicillin, and 100 μg/ml streptomycin (Sigma-Aldrich, Burlington, MA, United States) at 37°C in a humidified atmosphere of 5% CO2. NS3/4A-HCC36 cell lines by transfection of recombinant lentivirus consist package of plasmid: pCMVΔR8.91 (RNAi core, Academia Sinica, Taipei, Taiwan) and pMD.G (RNAi core, Academia Sinica, Taipei, Taiwan) and target plasmid: pLKO_AS3 NS3/4A, respectively; after selection by 1, 3, and 5 μg/ml puromycin dihydrochloride (P8833, Sigma-Aldrich, Burlington, MA, United States), the selection medium was replaced every 2 days for 1 week. Female BALB/c nude mice (6–8 weeks old) were obtained from the National Laboratory Animal Center, Taipei, Taiwan. All animal experiments were performed in accordance with institutional guidelines and approved by the Animal Care and Use Committee of Kaohsiung Medical University.