Reconstructing blood stem cell regulatory network models from single-cell molecular profiles

The aim of this study was to validate networks for haematopoietic stem and progenitor cell networks, using ChIP-Seq data to assess and confirm the genomic binding sites of important haematopoietic transcription factors in a haematopoietic cell line. Overall design: HoxB8-FL cells were fixed with either 1% or 0.4% formaldehyde for 10 mins. Chromatin was isolated, sonicated for 7 mins (30 sec on, 30 sec off), and specific antibodies were used to pull down the transcription factors of interest after a pre-clearing step. Chromatin was washed, de-crosslinked, amplified, size selected by gel purification and sequenced.