*4.9. Cell Cycle Analysis*

The proportion of cells in the Sub-G1 phase of the cell cycle, which indicate apoptotic hypodiploid cells were measured using flow cytometry. The experiments were carried out according to the method described by Fernando et al. (2018) [21]. Briefly, cells (2 × 10<sup>5</sup> cells mL−1) were incubated with di fferent concentrations of F5 for 24 h. The cells were then harvested, washed with PBS, and fixed in 70% ethanol followed by washing with EDTA (2 mM) in PBS. Then the cells were suspended in a PI solution containing RNase, and the cell cycle analysis was carried out using a FACSCalibur flow cytometer, USA.
