ELISA determination for nitrotyrosine in proteins of APAP-treated hepatocytes: time course and effect of various inhibitors of toxicity. Freshly isolated mouse hepatocytes were incubated with media alone or with APAP (1 mM) for 2 h. Subsequently, hepatocytes were washed to remove APAP (arrow) and incubated with media. To some incubations, cyclosporine A (10 μM) (CSP), N-acetylcysteine (1 mM) (NAC), SAIT (1 mM), or 7-nitroindazole (10 μM) (N7) was added to the pretreated cells for the remaining5hof incubation. Control cells were reincubated with media alone. At the indicated time, aliquots were taken, and 3-nitrotyrosine was analyzed using a commercial ELISA kit. Samples (n = 3 from separate mice) which significantly increased from the 2 h wash are indicated by * (p ≤ 0.05).