3.4. Apoptosis assay

The cells (400,000) were treated with vehicle, curcumin, docetaxel or combination of curcumin and docetaxel and after 48 hours of treatment, the apoptosis was detected by APC annexin V apoptosis detection kit with PI (Biolegend, San Diego, CA). Briefly, the cells were washed once with PBS, pellet was suspended in 100 mL of Annexin V Binding Buffer followed by incubation with annexin V (conjugated with APC) and propidium iodide (PI) in the dark for 15 minutes. The frequency of apoptotic cells was analyzed using a Guava easyCyte HT (EMD Millipore, Billarica, MA) Flow cytometer.