Casticin

The HCC MHCC97 cell line was purchased from Shanghai Xiangf Biotechnology Co., Ltd. (Shanghai, China). The MHCC97 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials Co., Ltd, Hangzhou, China), 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen Life Technologies, Carlsbad, CA, USA), and incubated in an atmosphere of 5% CO₂ at 37°C. Casticin was purchased from Chengdu Biopurify Phytochemicals Ltd. (Chengdu, China, dissolved in dimethyl sulfoxide (DMSO) as a 10-mmol/l stock solution, and diluted in a medium to the indicated concentration. MTT and lithium chloride were purchased from Sigma-Aldrich (St. Louis, MO, USA). Trypsin and DMSO were purchased from Amersco Company (Solon, OH, USA). Mouse anti-human β-catenin, cyclin D1, β-actin antibodies and horseradish peroxidase-conjugated rabbit anti-mouse secondary antibody were all purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

We purchased casticin from Chengdu Bio-purify Phyto-chemicals, Ltd. (Chengdu, China) and dissolved it in dimethyl sulfoxide (DMSO). The chemical inhibitor BYL719 was purchased from Selleckchem (Houston, TX, USA).