Fig. 2

AAE induces apoptosis and regulates pro-apoptotic proteins in HCT116 colon cancer cells. a Cell apoptosis observed using Hoechst 33342 staining. HCT116 were treated with AAE (0, 30, 40, and 60 ìg/ml) for 24 h. Fluorescence was detected using a fluorescence microscope. Arrows indicate apoptotic bodies, which were DNA fragments produced when apoptosis occurred. b Apoptotic effects of different concentration AAE were evaluated by Muse™ Annexin V and Dead Cell Assay Kit. HCT116 were treated with AAE (0, 30, 40, and 60 ìg/ml) for 24 h. Data analyzed by flow cytometry. c HCT116 were treated with 0, 30, 40, and 60 ìg/ml of AAE 24 h, caspase 3/7 activity was analyzed using a Muse™ Caspase-3/7 kit, as described in Materials and Methods. d Cells were treated with the indicated concentrations of AAE for 24 h. The expression of PTEN, PDK1, Akt (Thr308 and Ser473), MDM2, p53, PUMA,Bcl-2, pro-caspase3 and the activation of Bax, Bak and cleaved PARP were analyzed by western blot analysis