Genome wide glucocorticoid receptor (GR) binding sites upon cortisol treatment in human CD8 T cells
To map genome-wide chromatin occupancy of the human glucocorticoid receptor (GR; encoded by NR3C1) in primary activated human CD8? T cells, ChIP–seq was performed following physiological cortisol stimulation. CD8? T cells were isolated from healthy adult donors by immunomagnetic negative selection, activated in vitro with plate-bound anti-CD3 and anti-CD28, expanded in IL-2, and then re-stimulated with anti-CD3/anti-CD28 in the presence of hydrocortisone (cortisol; 100 nM) for 48 hours or vehicle control. GR-bound chromatin was immunoprecipitated and profiled by high-throughput sequencing to identify cortisol-induced GR binding sites and infer direct GR target loci in human CD8? T cells.