Global expression analysis of the transcriptome from bone marrow cells from wild type (WT) mice and mice with S1P-ablation in the Osterix-lineage (Cko)

Comparison of the transcriptome of non-RBC bone marrow cells from WT mice with non-RBC bone marrow cells from mice with S1P-ablation in the Osterix-lineage (Cko). Forelimbs and hindlimbs from mice were crushed in a mortar and pestle in the presence of phosphate buffered saline (PBS) and strained through a 70 micron filter and filtrate stored on ice. Crushed bones were digested with collagenase at 37 degree celcius for 15 min, strained through a 70 micron filter and pooled with the filtrate from above. The filtrate/cells were washed with cold PBS, and the RBCs lysed using an RBC lysis buffer (Sigma, R 7757) for 5 min. Following washings with cold PBS to remove lysed cells and hemoglobin, cells were suspended in 0.1% bovine serum albumin and processed by the 10x Genomics microfluidic platform (www.10xgenomics.com).