Beyort 3 cdna synthesis kit

Total RNA in cells was extracted by Trizol (KGA1202, KeyGEN Biotech, China), and then the synthesis of cDNA was operated through reverse transcription by the BeyoRT III cDNA Synthesis Kit (D7178, Beyotime, China). Next, cDNA and primers for detecting mRNA, as well as SYBR Green Supermix (A46113, Applied Biosystems, USA) for labeling, were added to a Fast7500 real-time PCR system (ABI, USA) to conduct the qPCR, and the thermal cycling program included predenaturation at 95°C for 2 min, 40 cycles of 95°C for 5 s and 60°C for 30 s. Following these, the internal reference gene GAPDH was utilized to calculate the mRNA level of the genes listed in Table 1 according to the 2^−ΔΔCt formula [24 (link)].

Mogroside V (MV, HPLC > 98%, Figure S1) was obtained from Nanjing Plant Origin Biological Technology Co., Ltd. (Nanjing, China; CAS:88901-36-4). Dorsomorphin (Compound C) was supplied by MedChemExpress (purity: 99.65%, Jersey, USA; CAS: 866405-64-3). Palmitic acid, oleic acid, and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were provided by Sigma (St. Louis, MO, USA). Triglyceride (TG), nonesterified fatty acid (NEFA), total cholesterol (T-CHO), aspartate aminotransferase (AST), and alanine transaminase (ALT) assay kits were obtained from Jiancheng Bioengineering Institute (Nanjing, China). Insulin ELISA kit was from Shanghai Hengyuan Biological Technology Co., Ltd (Shanghai, China). AMPK and p-AMPK (phospho-Thr172) antibodies were purchased from Santa Cruz (Dallas, USA); PPAR-γ, PPAR-α, FASN, CPT-1A, and α-tubulin antibodies were supplied by Proteintech (Chicago, USA); SREBP-1 antibody was purchased from Novus Biologicals (Littleton, Colorado, USA). TRIzol reagent was purchased from Invitrogen (Carlsbad, USA); BeyoRT™ III cDNA Synthesis Kit and BeyoFast™ SYBR Green qPCR Mix were purchased from Beyotime Institute of Biotechnology (Jiangsu, China).