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2 protocols using assay kits for cell counting kit 8

1

Endothelial Function Evaluation Protocol

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As-IV were obtained from Nanjing Jingzhu Biotechnology Co. Ltd. (Nanjing, China). Assay kits for Cell Counting Kit-8, NO nitrate reductase, dihydroethidium (DHE), GSH-px, MDA, SOD, JC-1, DAF-FM diacetate (DAF-FM DA) and MitoSOX were obtained from Beyotime Biotechnology (Shanghai, China). Calpain-1, SIRT1 and β-actin were obtained from Proteintech (Wuhan, China). Ser1177 eNOS phosphorylation, total endothelial nitric oxide (eNOS), Thr172 AMPK phosphorylation, total 5’ AMP activated protein kinase (t-AMPK), vascular cell adhesion molecule 1 (VCAM-1) and intracellular adhesion molecule 1 (ICAM-1) were obtained from ABclonal (Wuhan, China). Phenylephrine (PE) and acetylcholine (Ach) were obtained from Sigma Aldrich (Shanghai, China). MDL-28170, SRT1720 and L-NAME were obtained from Selleck (Houston, TX, United States).
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2

Ginsenoside Rg1 Attenuates Endothelial Dysfunction

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Ginsenoside Rg1 was obtained from Nanjin Jingzhu Biotechnology Company. Assay kits for Cell Counting Kit-8, NO nitrate reductase, dihydroethidium (DHE), MDA, JC-1, and MitoSOX were obtained from Beyotime Biotechnology (Shanghai, China). Antibodies against calpain-1 and β-actin were obtained from Proteintech (Wuhan, China). Antibodies against Drp1, Mfn2, PP2A, Ser1177 eNOS phosphorylation and endothelial nitric oxide (eNOS) were obtained from ABclonal (Wuhan, China). Phenylephrine (PE), acetylcholine (Ach), polyethylene glycol-catalase (PEG-Cat), 5-hydroxydecanoate (5-HD) and okadaic acid (OKA) were obtained from Sigma Aldrich (Shanghai, China). MDL-28170, L-NAME, acetylcysteine (N-acetyl-l-cysteine, NAC), TEMPOL and rotenone were obtained from Selleck (Houston, TX, USA).
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