2.14. Apoptosis Determination by Caspase-3 Activity Assay

The epidermis was incubated in 100 μL PBS containing 2.5 μM Ac-DEVD-AMC as a substrate, with 0.02% Triton X-100 and 10 mM DTT, at 37°C in a 96-well plate [43]. Fluorescence of the released coumarin derivative was measured at 390/435 nm, using a Fluostar-BMG spectrofluorometer (Offenburg, Germany). Caspase-3 activity was calculated over 40 min in a linear range from the fluorescence versus time slope. Results were normalized relative to the control group.