title: GSM8726410: CD117 cells, ei, Mouse 1,  H3K4me1 RELACS-ChIP; Mus musculus; ChIP-Seq
library_construction_protocol: For SPI1, GFI1 and LSD1 ChIP-seq, 10 million cells (per ChIP) were fixed with 1.5mM DSG and 1% formaldehyde before quenching with 125 mM glycine. After cell lysis and nuclei extraction, chromatin was sheared using a Covaris S220 to generate 100-500 bp fragments. Samples were immunoprecipitated with 5μg antibody either against GFI1 (R&D Systems, AF3540), SPI1 (Invitrogen, PA5-17505) or LSD1 (Schuele Laboratory, 20752). For H3K4me1 RELACS ChIP-seq, CD117+ bone marrow cells from LSD1 ei and LSD1 ei ctrl mice were isolated using CD117 MicroBeads (Miltenyi Biotec, 130097146) and LS columns (Miltenyi Biotec, 130042401) according to the manufacturer's instructions. 1-3 million CD117+ cells from Lsd1 ei and Lsd1 ei ctrl mice were fixed with 1% formaldehyde before quenching with 125 mM glycine and snap freezing in liquid nitrogen. Nuclei extraction, chromatin preparation and immunoprecipitation were performed as described in Arrigoni et. al (Communications Biology, 2018). For immunoprecipitation an antibody against H3K4me1 (Diagenode, C15410194) was used. For SPI1, GFI1 and LSD1 ChIP-seq,  Libraries were generated using an NEBNext Ultra II DNA Library Prep Kit (New England Biolabs, E7645). For H3K4me1 RELACS ChIP-seq, library preparation was performed as described in Arrigoni et. al (Communications Biology, 2018).