2.5. Real-time PCR
Total RNA was isolated from the liver using an RNase free DNase and
RNeasy Kit (Qiagen, Valencia, CA). cDNA was prepared using High Capacity
cDNA Reverse Transcription Kits (Applied Biosystems) and was subjected
to quantitative PCR using THUNDERBIRD SYBR qPCR Mix (Toyobo Co., Ltd.,
Osaka, Japan). The data were normalized according to the mRNA expression
levels of β-actin and TATA box-binding protein (Tbp). The primer
sequences are shown in Supplementary Table 1.