Bone Marrow - scRNA-sequencing

The goal of this experiment was to address when during development, deletion of the locus control region 1 (LCR1), impact the development of ILC2 in the bone marrow. In order to determine consequences of LCR1 deletion on ILC2 development, 8-12 weeks old WT and LCR1-/- C57BL/6J mice were sacrified and BM cells were extracted by crushing bones. Following BM cell extraction, ACK lysis for performed to remove red blood cell. Cells were filtered through a 70uM strainer and sequentially with viability dye and flow cytometry antibodies (in FACS buffer). Cells were then filtered again and sorted using a FACS ARIA II (BD). Viable, CD45+, Lineage-, CD127+ cells were sorted with a purity >90% then prepared for 10X Genomics scRNAsequencing experiment. scRNAseq experiment was performed following manufacturer's recommendations.