GSM8376424: m_WT_2; Mus musculus; RNA-Seq
Bone marrow cells were isolated by crushing long bones in 2% FCS using a mortar and then filtering the suspension through a 40 µm filter. Following centrifugation, the resulting pellet was resuspended in ACK lysis buffer for 30-40 seconds to lyse red blood cells, and the lysis was subsequently stopped with a 2% FCS solution. After centrifuging the solution, cells were stained for specific cell markers using fluorophore-conjugated antibodies for 30 minutes at +4. For RNAseq analysis, endothelial cells (ECs) from the bone marrow of mice were isolated directly into the lysis buffer of the RNeasy Plus Micro Kit ECs from the BM were defined as CD45-, Ter119- and CD31+, Sca1+. RNA was isolated according to manufacturer's instructions, and SmartSeq2 sequencing was performed. Illumina DNA Tagmentation followed by mRNA enrichment by poly-dT