Liensinine

Cycloheximide, 3-methyladenine, hydroxychloroquine, Spautin-1, orlistat, BEZ235, Cerulenin, and hydrogen peroxide were purchased from Sigma-Aldrich Chemie GmbH (Germany). Liensinine was obtained from ChemFaces (China) and bortezomib, carfilzomib, and palbociclib from Selleck Chemicals (USA).

Liensinine and nuciferine (≥98% by HPLC) were purchased from ChemFaces (Wuhan, Hubei, China) and dissolved with dimethyl sulfoxide (DMSO) and ethanol, respectively. Dulbecco's modified Eagle medium (DMEM), minimum essential medium alpha (α-MEM), Dulbecco's phosphate-buffered saline (PBS), fetal bovine serum (FBS), and antibiotic-antimycotic mixture containing 100 U/ml penicillin and 100 μl/ml streptomycin were obtained from Gibco BRL (Grand Island, NY). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), histopaque-1083, and DMSO were purchased from Sigma-Aldrich (St. Louis, MO). Anti-human Bcl-2, procaspase-3, caspase-3, poly(ADP-ribose) polymerase (PARP), and RANKL antibodies were obtained from Cell Signaling Technology (Beverly, MA). Anti-human Bax and horseradish peroxidase- (HRP-) conjugated anti-rabbit second antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Recombinant mouse soluble RANKL (sRANKL) and M-CSF were purchased from R&D system (Minneapolis, MN). All reagents used in this study were of analytical grade.