GSM9192815: Multiome, HSC, ATAC; Mus musculus; ATAC-seq
Bone marrow was isolated from the sternum, femur, tibia, and pelvis by crushing the bones in FACS buffer (PBS 5% FCS 2 mM EDTA) and filtering through a 50 µm strainer. WBC counts were measured using a haematology analyser. Cells were enriched for c-Kit expression using magnetic beads. After centrifugation, cells were resuspended at 1 million WBCs/µl, incubated with beads on ice, washed, and passed through a LS column. c-Kit+ cells were eluted for downstream processing. Cells were blocked, stained with a 2× antibody cocktail (FACS + CITE-seq), and resuspended in FACS buffer containing 7AAD. Long-term HSCs (LSK CD48− CD150+ CD34−) were FACS-sorted and processed following the DOGMA-seq protocol:: https://static-content.springer.com/esm/art%3A10.1038%2Fs41587-021-00927-2/MediaObjects/41587_2021_927_MOESM1_ESM.pdf ATAC and GEX libraries were prepared following the 10X Genomics protocol (CG000338 Rev F), with modifications based on the DOGMA-seq protocol to enable ADT library generation.