Metabolic changes during human adipocyte differentiation promote stable DNA hydroxymethylation and binding of acetylated NEIL1 to adipogenic enhancers [450k array]

In the present study, we applied two whole-genome sequencing techniques (WGBS/oxBS and hMe-Seal) to detect 5hmC (and 5mC) changes during the differentiation of the human SGBS preadipocyte cell line to mature adipocytes. As technical and biological validation we performed BS and oxBS followed by 450k array analysis. RNA-seq data was performed in parallel to study transcriptional changes associated with differential hydroxymethylation. In human white adipose tissue (WAT) hydroxymethylation (hME-Seal) was characterized in comparison with histone modifications and acNEIL1 binding (ACT-seq). DNA and RNA of SGBS cells at different time points of differentiation (day 0, 1, 7, 11 (and 27)) were examined for DNA methylation and hydroxymethlyation and mRNA expression levels in two indepentent experiments (Exp.1 and 2). DNA, RNA and Nuclei of 3-5 white adipose tissue biopsies were examined for DNA hydroxymethylation, mRNA expresion levels and histone modifications and acNEIL1 binding.