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2 protocols using anisomycin

1

Chondrocyte Oxidative Stress Regulation

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Anisomycin, Collagenase-II, Safranin-O/Fast Green, and dimethylsulfoxide (DMSO) were obtained from Solarbio Science & Technology Co., Ltd. (Beijing, China). Primary antibodies of Collagenase II, Aggrecan, ADAMTS-5, and MMP-13 were gained from Abcam (Cambridge, MA, USA). Primary antibodies directed against P-JNK and JNK were procured from Cell Signaling Technology (MA, USA). Primary antibodies against Bcl-2 and GAPDH were procured from ProteinTech Group (Wuhan, China). Primary antibodies against cleaved caspase-3 and Bax were procured from Affinity Biosciences (Cincinnati, OH, USA). Fetal bovine serum (FBS) and Dulbecco’s modified Eagle’s medium (DMEM)/F12 were obtained from Gibco (Grand Island, NY, USA). Tert-Butyl hydroperoxide solution (TBHP) was procured from Sigma-Aldrich (St Louis, MO, USA). The secondary antibodies of Goat Anti-Mouse IgG, Goat Anti-Rabbit IgG, Alexa Fluor®488 and labeled Alexa Fluor®594 were obtained from bioWORLD (OH, USA). TRIzol reagent was purchased from Invitrogen (Carlsbad, CA, USA). The BCA protein assay kit was procured from Beyotime Biotechnology (Shanghai, China). Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Laboratories (Kumamoto, Japan).The In Situ Cell Death Detection Kit was purchased from Roche (San Francisco, CA, USA). Caspase-3 colorimetric assay kit was obtained from Keygen Biotech Co., Ltd. (Nanjing, China).
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2

Esculetin Modulates Apoptosis Pathways

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Esculetin was purchased from Yuanyebio (Shanghai, China). Anisomycin (Ani), and methylthiazolyldiphenyl-tetrazolium bromide (MTT) was purchased from Solarbio (Beijing, China). A LDH Detection Kit and a JC-1 Staining Kit were purchased from Beyotime (Shanghai, China). An Annexin V-FITC/PI Apoptosis Detection Kit was purchased from Key gen Biotech (Nanjing, China). DMEM was purchased from Sigma Aldrich (St. Louis, IL, USA). Fetal bovine serum was purchased from ExCell Bio (Taicang, China). Antibodies against Bax, Bcl-2, Caspase-1, and p-c-Jun were purchased from Proteintech (Chicago, MO, USA). Antibodies against cleaved-Caspase-3, and p-c-Fos were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against c-Fos, c-Jun, Caspase-3, JNK, and PARP were purchased from Beyotime (Shanghai, China). Anti-β-actin antibody was purchased from Solarbio (Beijing, China). Antibodies against cleaved-PARP, Cytochrome c (Cyt-c), GSDMD, IL-1β, NLRP3, and p-JNK were purchased from Abcam (Cambridge, UK).
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