Fig. 2. Synergistic cytotoxic effect of combination of melatonin and zileuton in human renal tubular cells. (A) HKC-8 cells were treated with RSL3 (1 µM), melatonin (0-2 mM), or zileuton (0-50 µM) for 12 h. Cell viability was measured using Alamar blue assay (n=3). (B) HKC-8 cells were treated with RSL3 (1 µM), melatonin (1 mM), and zileuton (5 µM) for 6 h. Alamar blue assay was performed to determine the combination effects of melatonin and zileuton (n=3). (C) HKC-8 cells were treated with RSL3 (1 µM), melatonin (1 mM), and zileuton (5 µM) for 6 h. The synergistic cytotoxic effect of melatonin and zileuton in human renal tubular cells was confirmed by SYTOX green staining. Scale bar=25 μm. (D) HKC-8 cells were treated with RSL3 (2 µM), melatonin (1 mM), and zileuton (5 µM) for 30 min and then stained with BODIPY C11 for 30 min. BODIPY C11-stained cells were analyzed with a confocal laser-scanning microscope (Olympus) at 488 and 568 nm. Scale bar=40 μm. Data are presented as means ± SD (*p<0.05, **p<0.01 and ***p<0.001).
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