RNA Isolation and Quantification Protocol

Total RNA was isolated from the liver and AT using RNeasy Mini plus and RNeasy Lipid Tissue Mini kits (Qiagen, Wetburg, Leusden, The Netherlands), respectively, according to the manufacturer's instructions. RNA integrity was determined by agarose gel electrophoresis. RNA quantity (OD-260) and quality (260/OD-280) were determined using an ND-1,000 spectrophotometer (NanoDrop Technologies, Rockland, DE). Total RNA (1 μg) was converted into cDNA using a Quantitect Reverse Transcription kit (Roche, Mannheim, Germany) according to the manufacturer's instructions. Real time PCR (RT-PCR) was performed using a 7900HT system (Applied Biosystems, Warrington, UK) as previously explained [45 (link)] by using Power SYBR Green Master Mix (Roche, Mannheim, Germany). Values were corrected using the housekeeping gene Cyclophillin A (Ppia). Primer sequences are available in Supplementary Table 1.