Figure 2.

DAP attenuated H₂O₂-induced apoptosis in rat pheochromocytoma PC12 cells. PC12 cells were pretreated with various concentrations of daphnetin for 2 h, then stimulated with H₂O₂ (200 µΜ) for 24 h. (A) Cell morphology was detected using an inverted microscope. (B) The apoptotic ratio was determined by Annexin V-FITC/PI assay. (C) Cleavage of PARP (89/116 kDa) and caspase 3 (17/19 kDa), and the level of pro-caspase 3 (35 kDa) were detected by western blotting (β-actin, 45 kDa). (D) Cells were stained with 4′,6-diamidino-2-phenylindole, and nuclear morphology was observed and photographed using a fluorescence microscope. H₂O₂, hydrogen peroxide; DAP, daphnetin; FITC, fluorescein isothiocyanate; PI, propidium iodide; PARP, poly ADP-ribose polymerase.