RNAseq of F1 WAT
Total RNA was isolated from adipose and hypothalamus tissues using the RNeasy Lipid Tissue Kit (QIAgen) and from liver using TRIzol (n = 32, 4 animals per sex/diet/cross cohort). RNA concentration was measured via NanoDrop and RNA quality/integrity was assessed with a BioAnalyzer (Agilent). RNA-Seq libraries were constructed using the RiboZero kit (Illumina) from total RNA samples with RIN scores >8.0. Libraries were checked for quality and concentration using the DNA 1000LabChip assay (Agilent) and quantitative PCR, according to manufacturers protocol. Libraries were sequenced at 2x100 paired end reads on an Illumina HiSeq 4000. After sequencing, reads were de-multiplexed and assigned to individual samples.