Supplemental Materials: Adrenomedullin 2/Intermedin Exerts Cardioprotective Effects by Regulating Cardiomyocyte Mitochondrial Function
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- Material Type
- 文書・図像類
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- Zhao, YunluSakurai, TakayukiKamiyoshi, AkikoTanaka, MegumuIchikawa-Shindo, YukaKawate, HisakaMatsuda, YorishigeZhang, YanGuo, QianqianLi, PeixuanHoshiyama, KenHayashi, MarinaLi, JiakeShindo, Takayuki
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- eng
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- 一般
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- 1.Transcriptome analysis [Characteristics] strain: C57BL/6, gender: male, tissue: heart [Treatment protocol] Mice underwent transverse aortic constriction (TAC) or SHAM surgery, one week post-surgery, mice were anaestethized and hearts were harvested and stored at -80°C. [Extraction protocol] RNA was extracted with Trizol according to the manufacturer's instructions. [Label protocol] The fragmented 5.5 μg of single-stranded cDNA were labeled by biotin according to manufacturer's protocol using GeneChip WT PLUS Reagent Kit . [Hybridization protocol] 2.3 μg of the fragmented and labeled ss-cDNA were hybridized on Clariom S Mouse Array for 16 h at 45°C and 60 rpm in GeneChip Hybridization Oven 645. Arrays were washed and stained in GeneChip Fluidics Station 450. [Scan protocol] Arrays were scanned using GeneChip Scanner 3000 7G. [Data processing] The data were analyzed using Transcriptome Analysis Console (4.0) with default analysis settings and global scaling as normalization method.2.Metabolism analysis [Characteristics] strain: C57BL/6, gender: male, tissue: heart [Treatment protocol] Mice underwent transverse aortic constriction (TAC) or SHAM surgery, one week post-surgery, mice were anaestethized and hearts were harvested and stored at -80°C. [Extraction protocol] 20 mg of the heart tissue was homogenized with 1 mL 80% methanol (138-14521, Fuji Film, Tokyo, Japan), spun, and mixed at 4°C for 30 min; the supernatant was collected for analysis after centrifugation at 10,000 g for 5 min at 4°C. [Detection protocol] The extracts were purified using a CaptivaND Lipid filter plate (Agilent, Santa Clara, CA, USA) according to the manufacturer’s instructions, dried using a vacuum evaporator, resuspended in distilled water, and used for the detection of metabolites by LC-MS/MS analysis. The relative levels of metabolites in the central metabolic pathways were determined using the Method Package for Primary Metabolites (Shimadzu, Kyoto, Japan) with a Discovery HS F5-3 column (Sigma-Aldrich, Burlington, USA), according to the manufacturer’s instructions.JSPS KAKENHI Grant number: 21H02669The Otsuka Toshimi Scholarship Foundation(2021-2024).
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- 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）