*4.10. Reverse Transcription Polymerase Chain Reaction (RT-PCR) Analysis*

Total RNA was extracted from homogenized liver tissue by using TRIZOL reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. Total RNA (1 μg) was reversetranscribed into first-strand cDNA by using 200 units of MMLV-RT (Promega) in a total volume of 20 μL. For real-time PCR, a SYBR system with self-designed primers and 12.5 ng cDNA was used. The self-designed primers were as follows: CYP1A1 forward: GGTTCTGGATACCCAGCTGAC; reverse: TGTGGCCCTTCTCAAATGTCC, CYP1A2 forward: GCTGTGGACTTCTTTCCGGT; reverse: TGTCCTGGATACTGTTCTTGTTGA, CYP2C6 forward: TCCTGCTGAAGTGTCCAGAA; reverse: TGCAAGGGCTGCGATGTTT, CYP2C11 forward: TGAAGGACATCGGCCAATCA; reverse: CCCA TGCAACACCACAAAGG, CYP2D1 forward: ACCCATGGCTTCTTTGCTTTTC; reverse: GTCCTTGC TCCCGTACCAC, CYP3A1 forward: CTCAAGGAGATGTTCCCTGTCA; reverse: CAGGTTTGCCTT TCTCTTGCC, CYP3A2 forward: CCATCCACATCTGGTGGTCT; and reverse: TCAAAGGACGAG GACATGGTT. Amplification using 40 cycles of 2 steps (95 ◦C for 15 s and 60 ◦C for 1 min) was performed on an ABI Prism 7900HT sequence detection system (Foster City, CA, USA).
