Thymic lobes were cleaned from fatty tissues and incubated with Liberase and DNaseI (Roche Diagnostics; 200μg/ml and 30μg/ml, respectively; 45min, 37ºC) to obtain a cell suspension which was subsequently filtered through a nylon mesh (100micro pore size, Sefar Nitex) to remove debris. TEC were sorted and then RNA was either isolated from single cells using the SMART-seq2 protocol. TEC for ChIP-seq and ATAC-seq were treated as detailed in Žuklys S et al. Nature Immunology 2016:17(10):1206-1215 RNA-seq libraries were prepared using the appropriate Illumina kit for SMART-seq2 or HiSeq2500 (ChIP-seq). Nextera adapters were used for single cell RNA-seq, ATAC-seq and ChIP-seq.