GSM2254765: MPP_ATAC-seq_rep1; Mus musculus; OTHER
Bone marrow cells were flushed from the long bones (tibias and femurs) of mice with phosphate buffered solution (PBS) without calcium or magnesium. For detection of LSK cells, whole bone marrow cells were incubated with PE anti-mouse Lineage Cocktail antibody,  FITC-conjugated antibody to Sca1 (Ly6A/E; D7) and allophycocyanin (APC)-conjugated antibody to c-Kit (ACK2). CD150 and CD48 were measured with the following antibodies: phycoerythrin (PE)-Cy7 conjugated antibody to CD48 (HM48-1), BV421-conjugated antibody to CD150 (TC15-12F12.2), All antibodies were purchased from Biolegend except for antibodies to Sca1 and c-Kit, which were purchased from eBioscience. Antibodies to lineage, Sca1, c-Kit, CD150, CD48 were diluted 1:100. Nonviable cells were excluded using the viability dye 7-AAD (50 ug ml−1). Cells were sorted with a FACSAria (Becton Dickinson) automated cell sorter. ATAC-seq libraries were prepared as described in Buenrostro et al (Buenrostro et al. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. 2013. Nature Methods 10, 1213-1218, doi:10.1038/nmeth.2688