Caspase-Glo-3/7 assays

The caspase-Glo-3/7 assay is a homogenous, luminescent assay that measures caspase-3 and caspase-7 activities. Briefly, a total of 2X10⁴ cells per well were plated in a 96-well plate. Caspase-Glo-3/7 reagent (Promega, Madison WI) containing 100 μl of blank reaction, negative control cells and treated cells in culture medium was added to each well of a white-walled 96-well plate. The blank reaction was used to measure background luminescence associated with the cell culture system and caspase-Glo-3/7 reagent. The plate was covered with the lid and gently mixed at 300 rpm for 30 s. The plate was incubated at room temperature for 1 h. After incubation, the luciferase activity was measured and normalized using the Dual-Glo luciferase assay system (Promega, Madison WI) in Glomax Multidetection equipment according to the manufacturer’s instructions. Cell apoptosis rate was calculated as the subtracted value for the blank reaction from experimental values X 100. The luminescence value corresponds to relative light units. Each assay was carried out three separate times.