Nox Activity Measurement

Nox activity in CFs was evaluated using enhanced lucigenin chemiluminescence. The CFs were sonicated and homogenized. The supernatant was obtained after centrifugation at 12,000g for 10 minutes at 4°C and debris removal. NADPH (100 μM) was added to the supernatant as a substrate to react with Nox and generate superoxide anions. A microplate reader (BioTek, VT) was used to measure the light emission produced by the reaction of lucigenin (5 μM) and superoxide anions once every minute for 10 minutes. The value representing the Nox activity was expressed as the mean light units per minute per milligram of protein.