2 7 dichlorofluorescein diacetate dcfda cellular ros detection assay kit

Wedelolactone was provided by Chengdu Pufei De Biotech Co., Ltd. (purity > 98.0%, Chengdu, China). Prednisone acetate was from Huazhong Pharmaceutical Co., Ltd. (Xiangyang, China). Doxorubicin hydrochloride for injection was obtained from KeyGen. Co. Ltd. (Nanjing, China). Lecithin (average molecular weight: 760 g/mol) was purchased from Shanghai Advanced Vehicle Technology Pharmaceutical Ltd (Shanghai, China). Solutol^® HS15 (molecular weight: 963 g/mol) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Acetic acid, acetonitrile, and methanol were Chromatographic Grade and were obtained from Tedia Company Inc. (Fairfield, USA). A 2′,7′-dichlorofluorescein diacetate (DCFDA)-cellular ROS detection assay kit was obtained from Abcam (Cambridge, UK). The other reagents are analytical.

Water-soluble progesterone (PG), anesthetic MS-222 and apocynin were obtained from Sigma (St. Louis, MO, USA). Collagenase (280 U/mg) and catalase (11,000 U/mg) were purchased from Wako (Osaka, Japan), hCG was from Teikoku Zoki (Tokyo, Japan). Fluorogenic caspase-3 substrate IV was purchased from Calbiochem (La Jolla, CA, USA). Hydrogen peroxide colorimetric/fluorometric assay kit was from BioVision (Milpitas, CA, USA). Polyclonal anti-MAPK and anti-pMAPK antibodies were from Cell Signaling (Beverly, MA, USA), biotinylated anti-rabbit IgG was from Vector Laboratories (Burlingame, CA, USA). The Streptavidin Biotin Complex Peroxidase Kit, protein assay CBB solution, SOD (5000 U/mg) and BHA were from Nacalai Tesque (Kyoto, Japan). 2′,7′-dichlorofluorescein diacetate (DCFDA) Cellular ROS Detection Assay Kit was obtained from Abcam (Cambridge, UK), EUK 134 and MITO-TEMPO were ordered from Santa Cruz (Santa Cruz Biotechnology, Dallas, TX, USA). Other chemicals were obtained from Wako and Nacalai Tesque.

DMEM medium, DMEM red-phenol-free medium, RPMI 1640 medium, RPMI 1640 red-phenol-free medium, fetal bovine serum (FBS), L-glutamine and penicillin-streptomycin were purchased from Gibco BRL (Cergy-Pontoise, France). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), N-acetyl-L-cysteine (NAC), 3-methyladenine (3-MA), anti-β-actin antibody, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolocarbocyanine iodide (JC-1) and cell death detection enzyme-linked immunosorbent assay^PLUS (ELISA) were obtained from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Beclin-1, Atg5 and LC3 antibodies were acquired from Cell Signaling Technology—Ozyme (Saint-Quentin-en-Yvelines). 2’,7’-dichlorofluorescein diacetate (DCFDA) cellular ROS detection assay kit and goat anti-rabbit IgG H&L horseradish peroxidase (HRP) secondary antibody were purchased from Abcam (Paris, France). LysoTracker, MitoTracker, rabbit anti-mouse IgG-IgM H&L HRP secondary antibody, TO-PRO-3, annexin V-FITC and propidium iodide (PI) were obtained from Invitrogen—Thermo Fisher Scientific (Villebon-sur-Yvette, France). Immobilon Western Chemiluminescent HRP Substrate was acquired from Merck (Lyon, France). Caspase-3/7 green reagent was purchased from Sartorius (Göttingen, Germany).