Extraction of RNA and synthesis of cDNA

Total RNA was isolated from pieces of liver tissues (<30 mg) using a TissueLyser LT bead mill (Qiagen) and peqGOLD Trifast reagent (Peqlab, Erlangen, Germany) according to manufacturers' instructions. RNA qualities and concentrations were assessed using a Nanodrop ND-1000 UV-Vis spectrophotometer. SuperScript VILO cDNA synthesis kit (Invitrogen) was used to reverse transcribe 1 μg RNA into cDNA using 2 μl 5x VILO reaction mix and 1 μl 10x SuperScript enzyme mix in a final volume of 10 μl. After completion and termination of the RT reaction, cDNA was diluted with 190 μl LowTE and stored at −20°C.