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Cells were cultured at 37 C in Dulbeccos modified Eagles medium (DMEM) + GlutaMax 1965 (Gibco) supplemented with 10% fetal bovine serum and 1% penicillinstreptomycin in a humidified 10% CO 2 incubator

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Excess iron is stored in hepatocytes or macrophages via ferritin, while systemic efflux is regulated by ferroportin (FPN) and hepcidin ( 3+ is reduced to Fe 2+ by STEAP3 and exported to the cytosol via DMT1

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Most studies have shown that the presence of HO-2 indicates a crucial role in male reproductive system and brain related diseases, although some studies have found HO-2 in the prevention of kidney injury and diabetes