Quantifying Cell Death by NAMPT Inhibitors

To determine the cell death induced by NAMPT inhibitors, malignant cells were stained with ANNEXIN-V (ANXN, eBioscience, BMS306FI/300) and 7-aminoactinomycin D (7AAD, Immunotech, A07704) as described by the manufacturer and analyzed using flow cytometry. Dead cells were identified as ANXN+7AAD+ /7AAD+ and early apoptotic cells as ANXN+ 7AAD-. Specific cell death induced by inhibitors was calculated using the following formula: percent of cell death induced by compound = [(S – C) / (100 – C)] × 100; where S = treated sample cell death and C = untreated sample cell death.