GSM8366586: CD11b+F4/80+ general macrophages_from metastatic bone marrow_repeat 2; Mus musculus; RNA-Seq
Bone marrow cells were flushed out from the mouse femur and tibia with PBS containing 2% of FBS. Red blood cells were removed with RBC lysis buffer.The cell suspension was filtered through a 70-μm strainer and stained with live dye and antibodies. General macrophages were sorted by F4/80 and CD11b from single live bone marrow cells; Niche-specific macrophages were sorted by VCAM1, CD163, and CCR3 from single live bone marrow cells. Total RNA were extracted using RNAeasy micro kit from Qiagen The integrity of total RNA samples were assessed on Bioanalyzer 2100 using RNA 6000 Pico chip (Agilent Technologies, CA).  Messenger RNA was enriched from these samples using QIAseq FastSelect - rRNA HMR Kit (Qiagen, CA), fragmented and converted to cDNA and Illumina sequencing library using the PrepX RNA-seq library preparation protocol on the Apollo 324TM NGS Library Prep System (Takara Bio, CA).