Sk hep 1

The human hepatocellular carcinoma cell lines, Bel-7402 (Bel) and Bel-7402/5-fluorouracil (Bel/Fu), were purchased from KeyGen Biotech Co., Ltd. HepG2, SK-Hep-1 were obtained from obtained from the tumor cell bank of the Chinese Academy of Medical Science. The cells were cultured in RPMI-1640 medium (Gibco-BRL, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco-BRL, Carlsbad, CA, USA), 100 μg/ml penicillin and 100 μg/ml streptomycin at 37°C in a 5% CO₂ atmosphere. The Bel/Fu cells were maintained in medium containing 20 μg/ml 5-Fu. YAP specific siRNAs and full-length YAP complementary DNA were prepared and transfected as described previously [19 (link), 41 (link)].

HepG2, Sk-Hep-1, Huh-7, Hep3B and L-02 cells were purchased from Nanjing Key Gen Biotech Co. Ltd. (Nanjing, China). The cytotoxicity of test compounds was determined using an MTT assay. Briefly, the cell lines were incubated at 37 °C in a humidified 5% CO₂ incubator for 24 h in 96-microwell plates. After medium removal, 100 μL of culture medium with 0.1% DMSO containing test compounds at different concentrations was added to each well and incubated at 37 °C for another 72 h. The MTT (5 mg/mL in PBS) was added and incubated for another 4 h, the optical density was detected with a microplate reader at 490 nm. The IC₅₀ values were calculated according to the dose-dependent curves. All the experiments were repeated in at least three independent experiments.