RNA extraction and gene expression analysis

Total RNA was extracted from 100 mg liver sections in 1.3 mL TRIzol (Life Technologies, Carlsbad, California), with an additional phenol:chloroform extraction, as previously described (Nault et al., 2015b). RNA was stored in RNA storage solution (Life Technologies) and assessed for purity by Nanodrop (A₂₆₀/A₂₈₀ ratio; Thermo Scientific). Synthesis of cDNA was performed using 1 µg of RNA, High Capacity cDNA Reverse Transcription kit (Applied Biosystems), and MyCycler thermal cycler (Bio-Rad). Quantitative real-time PCR (qPCR) was performed using SYBR Green master mix (Bio-Rad) and a CFX96 Real-Time PCR Detection System (Bio-Rad). The expression of each gene was normalized to the geometric mean of Actb, Gapdh, and Hprt and differential expression was calculated using the 2^−ΔΔCT method. Primer sequences are provided in Supplementary Table S1.