A Quantitative Method to Measure Low Levels of ROS in Nonphagocytic Cells by Using a Chemiluminescent Imaging System

Figure 3

Comparison of chemiluminescence with or without 4IPBA on PMA-induced ROS generation in HT-29 cells. HT-29 cells () were resuspended in luminol (50 μM in NaOH) and HRP (0.2 U/ml) with or without 4IPBA (180 μM) in a black-walled and glass-bottomed 96-well plate. HT-29 cells were then (a) treated with SOD (50 μg/ml) and/or CAT (1.5 kU/ml) or (b) pretreated with NAC (10 mM) or DPI (10 μM) for 1 h. Chemiluminescence was measured using a luminometer for 30 min after PMA (200 nM) stimulation (, ±SD).

(a)

(b)