Fig. 3

Bicyclol induced G1 cycle arrest in HepG2 cells on a dose- and time-dependent manner. a The phase distribution of HepG2 cells treated with various concentrations (0, 50, 100, 200 and 500 μM) of bicyclol for 24 h was analyzed by flow cytometry. The HepG2 cells were plated in six-well plates and cultured until they reached 60 % confluence. The cells were incubated in serum-free RPMI 1640 culture medium for 24 h to be synchronized. Then the cells were treated with bicyclol for 24 h. The cell cycle distribution was determined by flow cytometry with the propidium iodide (PI) dye, and distribution of cells in G1, S, and G2 phases was calculated using the Cell Quest software. b The phase distribution of HepG2 cells treated with 200 μM bicyclol for 8, 16 and 24 h was analyzed by flow cytometry. The cells were treated as in (A). c The DNA distribution of cells treated with various concentrations of bicyclol for 24 h. d Dose-dependent effects of bicyclol on cell cycle-related proteins in HepG2 cells. The cells were disrupted after treatment with various concentrations (0, 50, 100, 200 and 500 μM) of bicyclol for 12 h. The proteins were collected, and cellular β-actin, cyclin D1, cyclin D3, cyclin E2, CDK2, CDK4, p21, p27 and p-Rb (Ser 807) were analyzed by western blotting. (*p < 0.05 versus PBS control)