Genome-wide STAT5 occupancy profiling in LPS induced CD127 expressing human monocytes

STAT5 functions in human inflammatory monocytes is largely unknown. We identified the anti-inflammatory function of CD127-STAT5 axis in human monocytes. To investigate the mechanism underlying STAT5-mediated anti-inflammatory regulation, thoroughly dissecting the STAT5 genomic distribution in human monocytes would be informative. Therefore, we performed anti-STAT5 ChIP-seq analyses for CD127high monocytes under LPS stimulation to globally characterize the STAT5 occupancy in CD127 expressing human monocytes. PBMC were isolated from buffy coat of healthy blood donors by Ficoll gradients, and CD14+ monocytes were isolated from PBMC with anti-CD14 magnetic beads (Miltenyi Biotec). CD14+ monocytes were treated with LPS for 6 h. Stimulated cells were firstly stained for surface CD127, and then were collected for FACS-based sorting of CD127high monocytes. Sorted cells were used to perform anti-STAT5 ChIP-seq experiment to globally characterize the STAT5 occupancy in CD127 expressing human monocytes.