Caspase activity kit

Caspase activities were measured with a caspase activity kit according to the manufacturer’s instructions (BioVision, Mountain View, CA, USA). Briefly, after different treatments, cells were washed with cold PBS, resuspended in lysis buffer, and left on ice for 15 min. The lysate was centrifuged at 16,000 g at 4 °C for 15 min. Activity of caspase 3 was measured with the use of the substrate peptide Ac-DEVDp-nitroanilide. We qualified the release of p-nitroanilide by determining the absorbance with a spectrophotometric plate reader at 405 nm. The fold increase in activity was calculated as the ratio between values obtained from treated samples versus those obtained from untreated controls.

Caspase activities were measured with a caspase activity kit according to the manufacturer’s instructions (BioVision, Mountain View, USA). The cells were washed with cold PBS, resuspended in lysis buffer, and left on ice for 15 min. The samples were then added to the reaction buffer containing Ac-DEVD-pNA, incubated for 2h at 37°C. The absorbance of yellow pNA, cleaved from its corresponding precursors, was measured using a spectrometer at 405nm. The fold increase in activity was calculated as the ratio between values obtained from the treated versus the untreated controls.