TRIzol RNA Isolation Reagent (Life Technologies, Gaithersburg, MD) was used for total RNA extraction from mouse liver [23 (link)]. Total RNA quantity was determined by an absorption spectrometer. The High Capacity cDNA Archive kit (Applied Biosystems, Foster City, CA, USA) was used for reverse transcription. Real-time RT-PCR was performed using TaqMan Universal PCR Master Mix (Applied Biosystems) according to the manufacturer’s instructions, and αSMA, collagen I, IL-1β, IL-6, IL-10, TNF-α, TGF-β and GAPDH were quantified using commercially available kits (TaqMan Gene Expression Assays Mm00725412_s1, Mm00801666_g1, Mm00434228_m1, Mm00446190_m1, Mm00439614_m1, Mm00443258_m1, Mm01178820_m1 and Mm03302249_g1, respectively; Applied Biosystems). These primer sets were designed to span one intron to allow identification of genomic contamination. Target gene results were calculated by delta Ct method (comparing target RNA expression to GAPDH). The reaction protocol consisted of the following cycles: 95°C for 15 min, 95°C for 15 sec and 60°C for 1 min for 50 cycles of PCR amplification on an Opticon 2 System (BioRad). All data were analyzed on an Option monitor 3 (BioRad).