Cell cycle analysis by flow cytometry

Cell cycle analyses was performed in Nalm6 cell line as described^29,33. A time course experiment was conducted on Nalm6 cells following treatment with quercetin (20 μM) for 8, 16 and 24 h. Besides Nalm6 cells were also treated with increasing concentrations of quercetin (0, 10, 50, 100, 250 μM) for 48 h. In both the experiments cells were seeded at 0.75 × 10⁵ cells/ml, cells were harvested after quercetin treatment, washed, fixed and incubated with RNase A. Cells were then stained with propidium iodide (50 μg/ml) and readings were acquired in flow cytometer (BD Biosciences FACS Calibur, USA). A minimum of 10,000 cells were acquired per sample and data was analyzed by using WinMDI 2.9 software. Experiments were repeated a minimum of two independent times and data is presented along with error bars.