*4.6. Real-Time PCR*

Total RNA was isolated from individual liver of each group for analysis of P38, ERK, JNK, IKK, c-jun, c-fos, P65, IRS1, PI3K, Akt2, and Glut2 using Trizol reagent (Invitrogen, Carlsbad, CA, USA) following the protocol provided by the manufacturer. Real-time quantitative PCR was performed by using SYBR Green Master mix and Rox reference dye according to the manufacturer's instructions. The cDNAs were obtained by the reverse transcription of RNA from rat liver. The mRNA levels of individual genes were normalized and calculated using the ΔΔCT method. The primers are listed in Table S1.
