Rna extraction reagent

Total RNA was extracted from cells using RNA extraction reagent (TRizol; Sigma). The total RNA isolated was reversed into cDNA using BioScript All-in-One cDNA Synthesis SuperMix (Biotool). Quantitative real-time PCR reactions were performed using the SYBR Premix Ex Taq (Tli RNaseH Plus) (Takara) in 7500 Real-Time PCR System (Applied Biosystems). For normalization, threshold cycles (Ct-values) were normalized to βactin/GAPDH within each sample to obtain sample-specific ΔCt values (ΔCt ¼ Ct gene of interest Ct βactin/GAPDH). The values of 2 -ΔΔCt were calculated to obtain fold expression levels. Primers for quantitative analyses of FGF21 gene (forward 5′-ATCGCTCCACTTTGACCCTG -3′, reverse 5′-GGGCTTCGGACTGGTAAACA -3′), GLP1-IgG4Fc gene (forward 5′-CCCCAAAACCCAAGGACACT -3′, reverse 5′-GCCATCCACGTACCAGTTGA -3′), srebp1c gene (forward 5′-CACTGTGACCTCGCAGATCC -3′, reverse 5′-ATAGGCAGCTTCTCCGCATC -3′), insulin gene (forward 5′-TCTCTACCTAGTGTGCGGGG -3′, reverse 5′-GCTGGTAGAGGGAGCAGATG -3′), β-actin gene (forward 5′-CCTGGCACCCAGCACAAT -3′, reverse 5′-GGGCCGGACTCGTCATAC -3′) and GAPDH gene (forward 5′-GGAGCGAGATCCCTCCAAAAT -3′, reverse 5′-GGCTGTTGTCATACTTCTCATGG -3′) were synthesized by TsingKe Company (Beijing).