4.5. Detection of Caspase 3/7 Activities

CellEvent™ Caspase-3/7 Green Flow Cytometry assay was performed according to the manufacturer’s instructions (Thermo Fisher, Vienna, Austria). Briefly, LNCaP C4-2, DU-145, RWPE-1 and HPrEC cells were plated at 50,000 cells per well in a 12-well plate, incubated overnight, and then treated with 50 mcg/mL of NaA-silane-PEG or NaA-silane-PEG-D2B for 96 h. After that time, cells were gently harvested by tripsin-EDTA and washed twice with cold PBS. Then, cells were incubated with CellEvent Caspase-3/7 Green Detection Reagent to a final concentration of 2 μM for 30 min. During the last 5 min of incubation, 1 µL of SYTOX AADvanced dead cell stain solution was added to each sample. Samples were run on a LSR II flow cytometer (BD Biosciences, San Diego, CA, USA) using FACSDiva software.