Gper sirna

Cryptotanshione (CPT) obtained from National Institutes for Food and Drug Control (Beijing, China) was prepared into a 10 mM stock solution dissolved by DMSO and stored at -20°C for later use. MTT (KeyGEN BioTECH, Jiangsu, China), DMEM-High Glucose medium (Hyclone company), Fetal bovine serum (Corning Cell Gro, Australia), 0.25% trypsin (Gibco, ThermoFisher Scinetific Inc, USA). Lipofectamine^TM 2000 Reagent was from Invitrogen Inc, USA. GPER specific agonist G-1 and antagonist G-15 were obtained from Cayman Chemical (Michigan, USA). The primary antibodies were listed below: cyclin D, cyclin B and cyclin A, Bax, Bcl-2, caspase-3, PI3K (p85) and GPER (Abcam, USA), p-AKT (Ser473, Cell Signaling Technology, USA). GPER siRNA, non-target siRNA and PI3K inhibitor LY294002 were purchased from Santa Cruz Biotechnology, USA.

Cryptotanshinone (CPT, purity>98%, molecular weight 296.36) was obtained from National Institutes for Food and Drug Control (Beijing, China). 2.964 mg CPT was dissolved in 1 ml dimethyl sulfoxide (DMSO, Sigma, USA) to make a 10 mM stock solution which was then added to the medium at the indicated concentrations. The methyl thiazolyl tetrazolium (MTT) was obtained from Keygen (Nanjing, China). RPMI-1640 medium and 0.05% trypsin were from Gibco (USA) and Fetal bovine serum was from Corning Cell Gro (Australia). Lipofectamine® 2000 was from Invitrogen (Grand Island, NY, USA). GPER specific agonist G-1 and antagonist G-15 were from Cayman Chemical (Michigan, USA). The following antibodies were used: GPER, PI3K(p85), cyclin A, cyclin B, cyclin D, CDK2 (Abcam, USA), phospho-Akt (p-AKT, Ser473, Cell signaling, Boston, MA, USA). GPER siRNA, non-target siRNA and PI3K inhibitor LY294002 were obtained from Santa Cruz Biotechnology (Texas USA). Fluorescein-Conjugated Goat anti-Rabbit IgG(H + L) was from ZSGB-BIO (Beijing, China) and DAPI was from Solarbio (Beijing, China).