GSM5268602: STAT5 ChIP-seq for CD127high monocytes; Homo sapiens; ChIP-Seq
Buffy coat from healthy blood donor was diluted by PBS, and PBMC were isolated by Ficol gradient. 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 fixed in 1% methanol-free formaldehyde. Then, cells were collected for FACS-based sorting of CD127high monocytes. Sorted cells were used to perform anti-STAT5 ChIP experiment by using the SimpleChIP enzymatic ChIP kit (Cell Signaling Technology) according to the manufacturer's instructions. ChIP-seq library construction with NEBNext® Ultra™ II DNA Library Prep kit (NEB, E7103S). Constructed DNA library was sent out for sequencing on an Illumina Hiseq X-ten platform for an average of 20 million unique reads per sample.