2.5. Apoptotic Cell Detection

3 × 10⁵ melanoma cells/well were cultured in complete medium for 18 h in 6-well plates. The medium was replaced by fresh medium supplemented with 1% FBS for the control condition or treated with Ocoxin 1:50, Vemurafenib 1 µM or the combination of both treatments for 48 h. Then, cells were washed with PBS, trypsinized, pelleted by centrifugation, washed 2 times with PBS, and double stained with the annexin V/IP apoptosis detection kit following manufacturer’s instructions (Thermo Fisher Scientific). Finally, apoptosis was determined by the Gallios cytometer (Beckman Coulter).