Illumina HiSeq 4000 paired end sequencing; Tissue-selective targeting redefines the mechanism of nuclear receptor Reverbα control over clock and metabolism - RNA-seq
Tissue-selective targeting redefines the mechanism of nuclear receptor Reverbα control over clock and metabolism - RNA-seq
Gonadal white adipose tissue (gWAT) or liver tissue was collected from adult male Reverbαflox/floxAdipoCre+/-, Reverbαflox/floxAlbCreERT2+/-, Reverbα-/- global knockout (KO) or Reverbα wildtype mice (littermates of global knockouts) at ZT8 and flash frozen (n=6-8). Frozen tissue was homogenised in TRIzol Reagent (Invitrogen) using Lysing Matrix D tubes (MP Biomedicals) and total RNA extracted according to the manufacturer's TRIzol protocol as far as the isopropanol phase, at which point samples were transferred to columns from the ReliaPrep RNA Miniprep Tissue Kit (Promega) to ensure high quality RNA.  The column was then washed, DNAse treated and RNA eluted as per Promega's protocol. Biological replicates were taken forward individually. Total RNA was submitted to the University of Manchester Genomic Technologies Core Facility (GTCF). Quality and integrity of the RNA samples were assessed using a 2200 TapeStation (Agilent Technologies).  Illumina TruSeq Stranded mRNA kit, according to the manufacturer's protocol
Experimental Factor: genotype
Reverbflox/floxAlbCreERT2+
Experimental Factor: organism part
liver