Genome analyzer 2 sequencing system

The total RNA was isolated from the perigonadal adipose tissue in F2 generation rats by using the E.Z.N.A. total RNA kit (Omega, Norcross, GA, USA). The quality and quantity were detected using a NanoDrop ND-1000 spectrophotometer (Thermo Scientific). RNA libraries were made according to the manufacturer's protocol of the TruSeq RNA sample preparation kit (Illumina, San Diego, CA, USA) and analyzed with the Genome Analyzer II sequencing system in the BGI Technology Services Co., Ltd., China. Differentially expressed genes were selected according to P < 0.05 or a fold change greater than 2.

Total RNA was extracted from LCM samples using a Qiagen RNeasy Lipid kit (Qiagen). The RNA quality and quantity were assessed using Agilent Bioanalyzer (Agilent) and NanoDrop ND-1000 spectrophotometer (Thermo Scientific), respectively. RNA libraries were prepared according to manufacturer’s protocol of TruSeq RNA sample preparation kit (Illumina, San Diego, CA, USA) and were sequenced with Genome Analyzer II sequencing system in the Genomics, Epigenomics and Sequencing Core at the University of Cincinnati. Analysis of gene expression was performed with our standard pipeline (Govindarajah et al. 2016 (link)). Differentially expressed genes of the HBF + BPA group were selected based on P < 0.05 and a fold-change greater than 2, when compared with the HBF-alone group. Functional enrichment analysis of differentially expressed genes was performed using the knowledge-based Ingenuity Pathways Analysis (IPA) (Qiagen, www.qiagen.com/ingenuity). RNA-seq data were deposited in the NCBI Gene Expression Omnibus database with accession number GSE73604.