4.4.1. Cell Death Profile

Treated and untreated yeast cells were washed and resuspended in 0.5 mL of PBS and labeled with propidium iodide (PI) and Annexin V, using the Annexin V-FITC apoptosis detection kit (Ebioscience, San Diego, CA, USA), according to the manufacturer′s instructions. Then, the cells were immediately analyzed on a FACS-Calibur flow cytometer (Becton-Dickinson, San Jose, CA, USA) by means of a 530/30 nm signal detector (FL1-H) for AV-FITC detection and 585/42 nm signal detector (FL2) for PI emission. Fluorescence intensity was acquired for 20,000 events in the closed region. Data were analyzed using FlowJo (Tree Star Inc., Ashland, DE, USA) and expressed as the percentage of cells in each population phenotype as compared to the control (untreated cells). Two independent experiments were performed (n = 4) [32].