Fig. 3
CBD induced apoptosis and G0-G1 cell cycle arrest by regulating the ER stress signaling. (A,B) Flow cytometric analysis of Annexin V-FITC/PI-stained cells. ES-2 cells were pre-incubated with 4-PBA, AM251 or NAC for 30 min at indicated concentrations. Cells were subsequently co-incubated with CBD at indicated concentrations. After 24 h, cells had been harvested and stained for analysis. Dot plots of total events are shown with frequencies of cells in each quadrant. (C,D) Cell cycle distribution was determined by flow cytometry. ES-2 cells were pre-incubated with 4-PBA or AM251 for 30 min at indicated concentrations. Cells were subsequently co-incubated with CBD at indicated concentrations. After 24 h, cells were harvested and fixed with 70% ethanol at 4 °C overnight, then, stained with PI. Each value indicates the mean ± SD of results obtained from three independent experiments. *p < 0.05, **p < 0.01, *** p < 0.001.
