Quantitative determination of gene expression via RT-PCR

Primary rat and human hepatocytes were treated with 10 μmol/L LEF or TER for 48 h. Total RNA was isolated with the TRIzol reagent (Life Technologies, CA, USA), and cDNA synthesis was performed using the PrimeScript RT Reagent Kit (Takara, Shiga, Japan). Quantitative analysis of the gene expression of several CYPs and transporters was conducted via real-time PCR using a Qiagen Rotor Gene Q instrument (Qiagen, Germany). β-Actin was used for internal normalization. All the primers (Table 1) were synthesized at Sangon Biotech (Shanghai) Co, Ltd.

F, forward primer; R, reverse primer; CYP, cytochrome P450 enzymes; BCRP, breast cancer resistant protein; BSEP, bile salt export pump; NTCP, sodium taurocholate co-transporting polypeptide; P-gp, p-glycoprotein.