Generation of Cellular Reactive Oxygen Species by Activation of the EP2 Receptor Contributes to Prostaglandin E2-Induced Cytotoxicity in Motor Neuron-Like NSC-34 Cells

Figure 7

Effects of NAC pretreatment on PGE2- and EP agonist-induced caspase-3 cleavage in differentiated NSC-34 cells. Differentiated NSC-34 cells were treated with vehicle (0.15% DMSO), 80 μM PGE2, 40 μM butaprost (an EP2 agonist), or 40 μM sulprostone (an EP1/3 agonist) in the presence or absence of 3 mM NAC for 8 h. Equal amounts of cell lysate (10 μg) were analyzed by Western blotting using anti-pro-caspase-3 antibody, anti-cleaved caspase-3 antibody, and also with anti-β-actin antibody as an internal control. The level of pro-caspase-3 (35 kDa) and cleaved caspase-3 (17 kDa) was assessed by densitometric analysis, and quantitative results were expressed as the ratio of the band intensity of pro-caspase-3 or cleaved caspase-3 to the band intensity of β-actin. Each value represents the for four different experiments. N.D.: not detected. , .