Schwann cell regulation by NR2F2 using Cut&Run

We focused on NR2F1 and NR2F2 (CoupTF1/CoupTF2) since they are expressed from neural crest through Schwann cell maturity, and found that knockdown of nuclear receptors Nr2f1 and Nr2f2 in primary Schwann cells downregulated genes such as Myelin Basic Protein (Mbp), Desert Hedgehog (Dhh), and N-Myc Downstream Regulated 1 (Ndrg1). In this study, we have elucidated a NR2F-regulated target gene network in Schwann cells, which revealed enrichment for non-myelinating Schwann cell genes. We used Cut&Run in S16 Schwann cells to show novel, genome-wide binding sites of NR2F1/2 and downstream transcription factors, YY1, SREBP1, Retinoid X Receptor (RXRG) and TEA-Domain factor (TEAD1). Our study elucidates the transcriptional cooperation that forms unique enhancer landscapes and the regulatory network that targets non-myelinating Schwann cells. Since nuclear receptor motifs are enriched in SOX10-bound enhancers in Schwann cells compared to oligodendrocytes (Lopez-Anido et al., 2015), we tested if NR2F1/2 proteins localize to SOX10-bound enhancers in S16 Schwann cells. We performed Cut&Run assays for NR2F2 (Skene and Henikoff, 2017). We also analyzed binding of NR2F1, but found that it is expressed at lower levels than NR2F2 and that the pattern of binding was less distinct compared to NR2F2. Finally, we performed ChIP-seq analysis for the enhancer mark, H3K27 acetylation (Rada-Iglesias et al., 2011), to identify those enhancers that are active in the S16 Schwann cell line.