Figure 4. The level of ROS increases after melatonin treatment. (A) The intracellular ROS production was measured with DCFH-DA after GBC-SD and NOZ cells were treated with or without 1 mM melatonin for 48 h. (B) Pre-treatment with 2 mM NAC for 1 hour significantly inhibited the production of ROS in GBC-SD and NOZ cells treated with 1 mM melatonin. (C, D) Pre-treatment with 2 mM NAC for 1 hour reversed the inhibition effects of melatonin in GBC-SD and NOZ cells. Three biological replicates were performed. Data are presented as mean ± SD. Mel, melatonin; ****P < 0.0001; ns, no significance.