4.11. Detection of Apoptosis by Flow Cytometry Assay

DU-145 cells were used for carrying out the in vitro apoptosis assay. Adequate cell number was sown into 6 well plates and allowed to adhere to growth media. After 24 h, a confluent monolayer was achieved which was separated into various groups; dimethyl sulfoxide control, sinigrin-treated, and colchicine-treated. The respective concentrations of the tested compounds were included in the confluent monolayer in the complete growth media and plates were incubated for 24 h, according to the regular MTT protocol. Cells were then washed thrice using phosphate-buffered saline, and later digested and centrifuged gradually. The detection of apoptotic cells was made by flow cytometry (Thermo Fisher Scientific) after staining with an Annexin V-FITC [54].