Cell Cycle Analysis of PC-3 Cancer Cells

A six-well plate was used, with 2 × 10⁵ PC-3 cells put into each well. The cells were grown in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 10% foetal bovine serum, and incubated for 24 h at 37 °C in a humid environment with 5% CO₂ in the air. The synthesised compound 5 was added to a brand-new incubation medium at its IC₅₀ concentration in DMSO (1% v/v). The cell plates underwent a 24 h incubation. The cells were fixed with 70% ice-cold ethanol after being rinsed twice with cold phosphate-buffered saline (PBS). Cells were centrifuged at 2000 rpm for 5 min after being washed with PBS at 37 °C for 30 min. Propidium iodide and DNA fluorochrome were used to stain the cells. Twenty minutes were spent incubating the plates at room temperature in the dark. Following that, the cells were examined using a FACS Calibre flow cytometer (Becton Dickinson, Heidelberg, Germany) ⁴⁸ (link)^,49 (link).