Fig. 3

Effect of compound 7d on cell cycle phase distribution of MCF-7 cells. The analysis of cells in different phases of the cell cycle was performed using propidium iodide (PI)-based assessment of cell cycle by flow cytometry. MCF-7 cells were seeded in 12-well plates and treated with different concentrations of 7d (20, 50, and 100 nM) for 48 h. The cells were then collected, washed with PBS and fixed with 70% ethanol overnight at −20 °C. After 24 h, the fixed cells were incubated in a PI-RNase solution at room temperature and 10,000 cells were analyzed using a FACS Calibur flow cytometer (BD Biosciences)