*4.7. RT-PCR*

Total RNA isolated from liver tissues was used to synthesize first-strand cDNA using a reverse transcriptase kit (DiaStartTM RT kit; SolGent, Daejeon, Korea) for RT-PCR and real-time PCR. As previously mentioned, the procedure for RT-PCR was performed [57]. Table 1 shows the primer sequences used to detect mRNA of IL-1β, IL-6, TNF-α, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH was used as a loading control. The PCR conditions included an initial denaturation at 94 ◦C for 5 min, followed by 30 cycles of 94 ◦C for 30 s, 58 ◦C for 30 s, and 72 ◦C for 30 s, and a final extension step at 72 ◦C for 10 min.

