Figure 3.

The PI3K/AKT signaling pathway was involved in the regulation of apoptosis and the cell cycle by Raddeanin A RA. (a) Immunoblot assay for the PI3K-AKT pathway. (b) Prior to treatment with RA (2 µM), the cells were treated with LY294002 (a PI3K inhibitor; 10 µM) for 1 h. The cells were incubated with annexin-V-FITC and propidium iodide (PI) and the apoptosis rate was analyzed via flow cytometry. (c) Protein levels of apoptotic proteins were determined via western blotting. β-actin was used as an internal control. (d) The mRNA and protein levels of cell cycle proteins were determined via RT-PCR and western blotting, respectively. β-actin was used as an internal control. The data are presented as the mean ± SD (n=3, ∗P<0.05, compared with the control).