Quantitative real-time PCR analysis
Total RNA from liver tissue samples were extracted by using Trizol
reagent. Single-strand cDNA was synthesized by reverse transcription of
total RNA using Primer Script RT reagent Kit (Takara, Japan). All
sequence of primers used for quantitative RNA analysis were listed in
Supplementary information. The mRNAs were amplified in Biosystems 7500
Real-time PCR reaction system using SYBR Premix Ex-Taq II Kit (Takara,
Japan) according to the manufactory’s instructions. The fold changes of
mRNA levels were analyzed by δδCt method.