Huh-7 and HepG2 Cell Culture Protocols

The cell lines Huh-7 (Japanese Collection of Research Bioresources, JCRB-0403) and HepG2 (American Type Culture Collection, ATCC-HB-8065) were cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, Gaithersburg, MD, USA) supplemented with 10% FBS (Hyclone, Thermo Fisher Scientific, Waltham, MA, USA), 100 U/ml penicillin, 100 mg/ml streptomycin at 37℃ in a humidified 5% CO₂ atmosphere. A stable Huh-7 cell line, which overexpresses FLAG-tagged 14-3-3ε, was previously established and maintained in DMEM with 10% FBS and 200 μg/ml G418^{36 (link)}. All pharmacological inhibitors were purchased from Sigma–Aldrich (St. Louis, MO, USA). Recombinant human DKK-1 was purchased from PeproTech (Rocky Hill, NJ, USA).