GSM7908857: long bone, HSC, day 14 post 5-FU, biological replicate 3; Mus musculus; RNA-Seq
after obtaining a single cell suspension from either spleen or bone marrow (long bones), cells were subjected to magnetic cell separation to enrich for lineage negative cells (CD11b-/Gr-1-/CD4-/CD8-/Ter119-/B220-). Subsequently, cells were stained with hematopoietic stem cell markers and finally 100 cells of the immunophenotype Lineage-/EPCR+/Sca-1+/CD48-/CD150+ (also termed long-term HSCs) were sorted into cell lysis buffer to extract RNA. After 100 cells were sorted, first strand cDNA were synthesized followed by generating 2nd strand by minorly modified method that was previously reported (Hiyashi et al. Nature Communication, 2018: 9). Using Nextera XT DNA sample Prep Kit (Illumina Inc., San Diego, CA), libraries were prepared by minorly modified method that was previously reported (Hiyashi et al. Nature Communication, 2018: 9) .