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. Overall design: 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.