Ns 398

PGE2 and NS398 were purchased from Cayman (Ann Arbor, USA); ONO-AE3-208 from ONO Pharmaceuticals, Osaka, Japan. Dr Peeyush K. Lala at the University of Western Ontario kindly provided us with all these chemicals.

The retinal ganglion cell line, RGC-5, from the American Type Culture Collection (ATCC, Manassas, VA, USA) was cultured in Dulbecco’s modified Eagle’s medium (DMEM; HyClone Laboratories, Red Bank, NJ, USA) containing 10% fetal bovine serum (FBS; Hyclone Laboratories) and 1% penicillin/streptomycin mixture (HyClone Laboratories) at 37°C in a humidified atmosphere of 5% of CO₂. The rhesus retinal vascular endothelial cell line, RF/6A (from the Cell Bank of the Chinese Academy of Sciences, Shanghai, China), was cultured in RPMI-1640 medium (Gibco, Grand Island, NY, USA) containing 10% FBS and 1% penicillin/streptomycin at 37°C in a humidified atmosphere of 5% CO₂. The culture medium was replaced with fresh medium every other day, and the cells were passaged every 2–3 days. The RGC-5 cells were incubated with succinate with or without pre-treatment with 10 µM of the ERK1/2 inhibitor, U0126 (Calbiochem, Gibbstown, NJ, USA), 10 µM of the JNK inhibitor, SP600125 (Calbiochem), or 50 µM of the COX-2 inhibitor, NS-398 (Cayman Chemical Co., Ann Arbor, MI, USA).