Cell Cycle Analysis of HCC Cells

This procedure was reported previously [31 (link)]. In brief, HCC cells were cultured in 6-well plates and treated with increased doses of UA for 24 h. Afterwards, the cells were harvested, and resuspended in 500 μL of cold PBS for 2 h at 4 °C. Following washes, the fixed cells were incubated in 1 mL of 0.1 % sodium citrate containing propidium iodide (PI) 0.05 mg and 50 μg RNase for 30 min at room temperature (RT), subjected to FACSCalibur flow cytometric analysis (FC500, Beckman Coulter, FL, USA). The proportion of cells within the G0/G1, S and G2/M phases was analyzed using the MultiCycle AV DNA Analysis software (Phoenix Flow Systems).