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
Dysbiosis, an imbalance in microbial composition, has been linked to a range of GI disorders, including inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), celiac disease, and colorectal cancer
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