Cell Cycle Analysis of Plant Essential Oils

Analysis of cell cycle progression was assessed by flow cytometry using a PI/RNase solution (Immunostep, Salamanca, Spain), according to the manufacturer’s protocol. Briefly, cells were treated with two different concentrations (0.7 or 0.8 µL/mL) of either M. cervina EO or L. pedunculata EO and further incubated for 24 h. After incubation, cells were detached and fixed in 70% ethanol for 1 h at 4 °C. Then, cells were stained with 500 µL of PI/RNase solution. Results were acquired using CellQuest software and analyzed to calculate the percentage of the cell population in subG0/G1, G0/G1, S, and G2/M cell cycle phases.