HuR knockdown in humanized and wildtype mice liver samples

HuR shRNA adenoviruses were delivered into WT or humanized mice intravenously at 2 × 109 pfu/mouse for both control virus and HuR shRNA virus . After seven days, liver tissue samples were harvested after a four hours fasting and stored immediately in liquid nitrogen till further analysis.The frozen liver tissue samples were homogenized in Trizol reagent (Invitrogen) using TissueLyser LT system (Qiagen). The isolated RNA was purified by MagMAX RNA Extraction Kit (ThermoFisher) and the construction of strand specific sequencing libraries using TruSeq Stranded Total RNA Prep kit (Illumina) and the sequencing was performed at NHLBI DNA Sequencing and Genomics Core using Illumina HiSeq 3000 paired-end sequencing platform. RNA sequencing profiles were generated for HuR konckdown in humanized and wild type mice. The humanized livers are generated by engrafting human hepatocytes in immunodeficient mice livers.