Figure 1: Effect of piceatannol on the proliferation (A), cell cycle (B), and apoptosis of AH109A hepatoma cells (C). (A) Piceatannol was dissolved in ethanol. The piceatannol solution was added to the culture medium at a final ethanol concentration of 0.2%. Piceatannol was suspended in a 0.3% carboxymethyl cellulose sodium salt (CMC) aqueous solution at concentrations of 0.25, 0.5, and 1 mg/mL. Piceatannol suspension or 0.3% CMC alone (vehicle control) (1 mL/100 g body weight) was intubated to male Donryu rats which had been fasted overnight, and blood was collected 2 h after oral administration. The proliferative activity was determined by [methyl-3H]thymidine incorporation method in vitro (A(a)) and ex vivo (A(b)). Each value represents the mean ± SEM of six wells. Values not sharing a common letter are significantly different at . (B) For cell cycle analysis, 3.0 × 105 cells of AH109A per well were seeded in the medium containing 0, 25, and 50 μM piceatannol and cultured for 24 h. Cells were then collected and washed twice with PBS(−). Thereafter, propidium iodide (PI) solution was added and cells were incubated for 30 min on ice. Cells at different cell cycle phases were then analyzed with a flow cytometer. Data are from representative experiment repeated three times with similar results (B(a)–(c)). (C) The effect of piceatannol on apoptosis in AH109A cells was assessed using Annexin V-FITC kit according to the manufacturer’s instructions. Briefly, 5 × 105 cells of AH109A per well were seeded in a 6-well plate and cultured in the medium containing 0, 50, and 100 μM piceatannol for 3 h. At the end of culture, cells were labeled with Annexin-V-FITC and analyzed with a flow cytometer. Data are from representative experiment repeated three times with similar results (C(a)–(c)).