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For cell lysis: The harvested cells were resuspended in buffer A (50 mM Tris-HCl, pH 8.0, 500 mM NaCl, 20% Glycerol, 1 mM tris(2-carboxyethyl)phosphine (TCEP), 0.1% IGEPAL CA-630, 0.5% (W/V) 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) and 10 mM Imidazole) containing EDTA-free protease inhibitor cocktail

In patients with thalassemia, the high levels of growth differentiation factor 15 (GDF15) and erythroferrin can also inhibit the production of hepcidin (55, 56)