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2 protocols using antibodies specific for β actin

1

Mitochondrial Dynamics and Oxidative Stress

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Dulbecco’s modified Eagle’s medium (DMEM), RPMI 1640 medium, fetal bovine serum, penicillin/streptomycin (10,000 units/mL penicillin and 10,000 µg/mL streptomycin), and 0.25% trypsin-EDTA were purchased from Gibco, Thermo Fisher Scientific, Inc. (USA). JC-1 dye was obtained from Invitrogen, Thermo Fisher Scientific, Inc. (USA). Nar, APAP, dorsomorphin (Dor), and brusatol (Bru) were obtained from MedChemExpress (USA) and dissolved in dimethyl sulfoxide (DMSO). A malondialdehyde (MDA) assay kit (TBA method), superoxide dismutase (SOD) assay kit (WST-1 method), reduced GSH assay kit, and glutamate dehydrogenase (GDH) test kit were obtained from Nanjing Jiancheng Bioengineering Institute (China). A reactive oxygen species (ROS) assay kit was obtained from Beyotime (China). A CCK8 Cell Proliferation and Cytotoxicity Assay Kit was obtained from Solarbio (China).
Antibodies specific for β-actin were purchased from Proteintech Group, Inc. (China). Antibodies specific for mitofusin protein 1 (Mfn1), optic atrophy type 1 (Opa1), phospho (p)-dynamin-related protein 1 (Drp1) (Ser616), total Drp1, AMPK, p-AMPK (Thr172), Nrf2, and GAPDH were purchased from Cell Signaling Technology (USA).
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2

Esculetin-Mediated Apoptosis Regulation

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Esculetin was obtained from Nanjin Jingzhu Biotechnology Company (purity>98%; Nanjing, China); 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Amresco (USA); The Annexin V-FITC apoptosis detection kit, DAPI and caspase-3/-9 were purchased from Beyotime Institute of Biotechnology; Antibodies specific for IGF-1 Bcl-2, Bax, PI3K, Akt, cytochrome c were purchased from abcam Technology; Antibodies specific for β-actin, GAPDH and VDAC were purchased from proteintech; IGF-1 and LY294002 were purchased from sigma company; RPMI 1640 medium and calf serum were purchased from Gibco.
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