Abi gene express 2.0 program

Total RNA was isolated from the cells, or dorsal tissue using an Easy Blue kit (Intron Biotechnology, Inc., Seoul, Korea) according to the manufacturer’s protocol. Total RNA was quantified using an Epoch micro-volume spectrophotometer system (BioTek Instruments, Inc., Winooski, VT, USA). cDNA was obtained using isolated total RNA (2 μg), d (T)16 primer, and Avian Myeloblastosis Virus reverse transcriptase with genomic DNA remover. The relative gene expression was quantified using RT-qPCR analysis (Real Time PCR System 7500; Applied Biosystems; Thermo Fisher Scientific, Inc., Waltham, MA, USA) with SYBR Premix Ex Taq. Fold changes of gene expression were calculated using the comparative quantification cycle (Cq) method. The Cq values of target genes were normalized to that of GAPDH using the ABI gene express 2.0 program (Applied Biosystems; Thermo Fisher Scientific, Inc., Waltham, MA, USA).

Total RNA was isolated from the liver tissues, adipose tissues, or cells using an Easy Blue kit (Intron Biotechnology, Inc., Seoul, Korea) according to the manufacturer’s protocol. Total RNA was quantified using an Epoch micro-volume spectrophotometer system (BioTek Instruments, Inc., Winooski, VT, USA). cDNA was obtained using isolated total RNA (2 μg), d(T)16 primer, and Avian Myeloblastosis Virus reverse transcriptase with genomic DNA remover. Relative gene expressions were quantified using RT-qPCR analysis (Real Time PCR System 7500; Applied Biosystems; Thermo Fisher Scientific, Inc., Waltham, MA, USA) with SYBR Premix Ex Taq. Fold changes in gene expression were calculated using the comparative quantification cycle method. The Cq values of target genes SREBP-1, PPARγ, C/EBPα, TNF-α, IL-6, IL-1β, MCP-1, leptin, adiponectin, FOXO1, SIRT1, AMPK, LKB1, and CaMKK were normalized to that of GAPDH using the ABI Gene Express 2.0 program (Applied Biosystems; Thermo Fisher Scientific, Inc., Waltham, MA, USA). The sequences of oligonucleotide primers are described in Table 1.