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Using the 2 CT method [ relative expression (fold change) of forkhead box O1 (FoxO1) (5-AGCTCAAACGCTAGCACCAT-3 and 5-GGTGGATACACCAGGGAATG-3), glucagon (5-GCCGAGCAAGGCGAGACT-3and 5-CATGTCTGCGCCCAAGTTC-3), insulin (5-CCTGCCCAGGCTTTTGTCA-3 and 5-GGTGCAGCACTGATCCACAATG-3), X-box binding protein 1 (XBP1) (5-GGTCTCAGAGGCAGAGTCCAAG-3 and 5-AGAGGCAACAGCGTCAGAATCC-3), Hspa5(BIP) (5-GAGGACAAGAAGGAGGATG-3 and 5-TTGGACGTGAGTTGGTTC-3), IL6 (5-GTCAACTCCATCTGCCCTTC-3 and 5-TGTGGGTGGTATCCTCTGTG-3), IL1 (5-GCCAACAAGTGGTATTCTCCA-3 and 5-TGCCGTCTTTCATCACACAG-3), HIF (5-TGGATGGCTTTGTTATGGTG-3 and 5-TGGTCACATGGATGGGTAAA-3), and GAPDH (5-TTAAGGGCATCCTGGGCTACACT-3 and 5-TTACTCCTTGGAGGCCATGTAGG-3) was normalized to -actin (5-GTCGTACCACTGGCATTGTG-3 and 5-CTCTCAGCTGTGGTGGTGAA-3) as the most suitable reference gene (expression level unaffected by the experimental treatment) and relative to the sham group as the calibrator

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It allows researchers to study how parts of human growth hormone can affect fat and energy metabolism, without the full hormones effects

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