Fas associated protein with death domain fadd

Human ovarian carcinoma cell lines A2780/CP70 and human immortalized ovarian surface epithelial cells (IOSE 364), were kind gifts from Dr. Bing-Hua Jiang at Thomas Jefferson University and Dr. Auersperg at the University of British Columbia, respectively. Cells were cultured in RPMI 1640 medium (Sigma, St. Louis, MO, USA) incorporating 10% fetal bovine serum (FBS) (Invitrogen, Grand Island, NY, USA). Cells were grown in a humidified incubator containing 5% CO₂ at 37 °C.
Reagents: Kaempferol was purchased from Sigma. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor^® 488 and propidium iodide (PI) was purchased from ThermoFisher Scientific (Waltham, MA, USA). Caspase-Glo^® 3/7 Assay Systems, Caspase-Glo^® 8 Assay Systems and Caspase-Glo^® 9 Assay Systems were purchased from Promega (Madison, WI, USA). Antibodies against p21 Waf1/Cip1 (p21), p-Cdc25C (Ser 216), Cdc25C, p-Cdc2 (Tyr 15), Cdc2, Cyclin B1, p53, death receptor 5 (DR5), Fas, and Fas-Associated protein with Death Domain (FADD) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibodies against phosphor-checkpoint kinase 2 (Chk2) (Thr68), Chk2, poly [ADP-ribose] polymerase 1 (PARP-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).