Fig. 3. Inhibition of ferroptosis by the combination of melatonin and zileuton via AKT/mTOR/NRF2 signaling pathway. (A) HKC-8 cells were treated with RSL3 (2 µM), melatonin (1 mM), and zileuton (5 µM) for 30 min and then stained with FerroOrange for 30 min. FerroOrange stained cells were analyzed with a confocal laser-scanning microscope (Olympus) at 488 and 568 nm. Scale bar=50 μm. (B) HKC-8 cells were treated with RSL3 (1 µM), melatonin (1 mM), and zileuton (5 µM) for 12 and 24 h. The expression levels of 4-HNE, GPX4, HO-1, p-AKT, p-mTOR, and Nrf2 were determined using western blotting analysis. Data are presented as means ± SD (***p<0.001).
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