Total RNA from both tissue and cells was isolated by using TRIzol reagent (CoWin Biosciences) according to the manufacturer's protocol. The perfused mouse liver tissues from normal 6-week-old C57BL/6 mice (n=5 males/group) and drug or PBS-treated 6-week-old C57BL/6 mice (n=5 males/group) were dissected out and homogenized in the TRIzol reagent. The recovered liver tissue and cells were lysed in TRIzol reagent. Total RNA was extracted and subjected to reverse transcription reactions with hexamer and M-MuLV reverse transcriptase (New England Biolabs, Inc.). The cDNA products were additionally diluted and used as PCR templates. The gene-specific PCR primers (Table SI) were designed using Primer3 Plus (http://www.bioinformatics.nl/cgi-bin/primer-3plus/primer3plus.cgi). TqPCR was performed using SYBR Green-based TqPCR analysis on a CFX-Connectunit system (Bio-Rad Laboratories, Inc.), as described previously (54 (link)). TqPCR reactions were performed in triplicate. GAPDH was used as the reference gene. Quantification of gene expression was performed using the 2−ΔΔCq method (55 (link)).