4.6. Gene Expression Analysis Using a Quantitative Polymerase Chain Reaction (qPCR)

Quantification of the gene expression of 3T3-L1 adipocytes and PWATs was performed using QuantStudio3 RT-PCR system (Applied Biosystems, Thermo Fisher Scientific, San Jose, CA, USA). Amplification was performed for each target gene using 5.5 µl cDNA, 1 µl each primer (10 µM, forward and reverse), 10 µl 2X TB green mix (TaKaRa Bio, Kusatsu, Shiga, Japan), and 2.5 µl deionized water. The expression levels of the target genes were normalized to β-actin. Table 1 shows the primer sequences used for qPCR.

Primer sequences for quantitative real-time PCR.

C/EBPα: CCAAT/enhancer-binding protein alpha. PPARγ: peroxisome proliferator-activated receptor gamma. SREBP1: sterol regulatory element-binding transcription factor 1.