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RNA isolation and quantitative RT-PCR Total RNA from cells was isolated with Trizol reagent (Qiagen GmbH, Hilden, Germany) and RT-PCR was conducted using a quantitative SYBR Green RT-PCR kit (Applied Biosystems, Warrington, UK) was used with a Step One Plus RT-PCR System (Applied Biosystems)

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Rather than utilizing a single pathway, this research kit pairs a highly robust growth hormone secretagogue receptor (GHS-R1a) agonist with a growth hormone-releasing hormone (GHRH) receptor agonist