Ferritin

Mouse plasma concentrations of ALT and iron were measured using an automatic analyzer (Hitachi High-Technologies Corporation, Tokyo, Japan). Hepcidin concentration was analyzed by liquid chromatography/electrospray ionization–tandem mass spectrometry. Ferritin (ALPCO, Salem, NH, USA) and BMPER (USCN Life Science Inc., Wuhan, China) were analyzed by enzyme-linked immunosorbent assay (ELISA).

Hematological parameters were measured by an automated hematology analyzer KX-21NV (Sysmex Corporation, Hyogo, Japan). The levels of cytokines and albumin present in serum were determined by using commercially available ELISA kits for human IL-6, mouse erythropoietin (EPO) (R&D Systems, Minneapolis, MN, USA), mouse serum amyloid A (Life Technologies Japan, Tokyo, Japan), mouse albumin (Shibayagi, Gunma, Japan), and ferritin (ALPCO Diagnostics, Salem, NH, USA). Serum iron level was determined by QuantiChrom Iron Assay Kit (BioAssay Systems, Hayward, CA, USA). Mouse interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and IL-6 were measured by Bio-Plex Pro cytokine assays according to the manufacturer’s instructions (Bio-Rad Laboratories, Hercules, CA, USA). The assays were performed using the Bio-Plex Pro II wash station with magnetic plate carrier, and cytokines were determined by the Bio-Plex 200 System (Bio-Rad Laboratories).