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2 protocols using ferrous sulfate heptahydrate

1

Cellular Responses to Iron Compounds

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Ferric sulfate and ferrous sulfate heptahydrate were purchased from Energy Chemical Technology Co., Ltd. (Shanghai, China). Ferric citrate was purchased from Sigma‐Aldrich (St. Louis, MO). Fetal bovine serum (FBS) was purchased from Invitrogen (Carlsbad, CA). Dulbecco's Modified Eagle's Medium (DMEM) was obtained from Sigma (Yuzhong District, Chongqing, China). Antibodies against p21 and C646, a p300/CREB‐binding protein (CBP) inhibitor, were purchased from Abcam Trading Co. Ltd. (Shanghai, China). N‐acetyl‐l‐cysteine (NAC), dihydroethidium (DHE), and a ROS Detection Kit were purchased from Beyotime Company (Jiangsu, China). Prussian Blue staining solution was acquired from Leagene Biological Technology Co. Ltd. (Beijing, China). Antibodies against p53, acetylated p53, CD86, and CD206 were from Cell Signaling Technology (Danvers, MA), and IL‐1, IL‐10, TGF‐β, and TNF‐α were purchased from GeneTex (San Antonio, TX). p21, p300, CBP, p65, phosphorylated p65, and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) were purchased from Abcam (Cambridge, MA). FITC‐CD86 and APC‐CD206 were purchased from Thermo Fisher Scientific (Waltham, MA).
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2

Antioxidant Capacity Evaluation Protocol

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Salicylic acid, 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), ABTS, ferrous sulfate heptahydrate (FeSO 4 •7H 2 O), L-ascorbic acid, cupric chloride dihydrate (CuCl 2 •2H 2 O), ammonium acetate (NH 4 Ac), 2,9-dimethyl-1,10-phenanthroline (Neocuproine, Nc), quercetin, D-(+)-glucose, butylated hydroxytoluene (BHT), butyl hydroxyanisole (BHA), tertiary butylhydroquinone (TBHQ) were purchased from Energy Chemical. 6-Hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) and β-carotene were purchased from TCI. DPPH was purchased from Alfa Aesar. Folin & Ciocalteu's phenol reagent (FC reagent), linoleic acid, aluminium chloride hexahydrate (AlCl 3 •6H 2 O) were purchased from Xiya Reagent. Sodium carbonate (Na 2 CO 3 ), methanol, ethanol, acetone, ethyl acetate, chloroform, hexane and dimethyl sulfoxide (DMSO) were purchased from Sinopharm. High-glucose Dulbecco's modified Eagle's medium (HG-DMEM) was purchased from Hyclone. Fetal bovine serum (FBS) was purchased from Gibco. 100× penicillin-streptomycin solution was purchased from Biosharp. Cell Counting Kit-8 (CCK-8) was purchased from Beyotime. TM3 mouse leydig cells were purchased from Procell.
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