GSM8529723: MDA-MB-231 cells cultured in endosteal bone containing only osteoblasts (OB) sample 2; Homo sapiens; RNA-Seq
At day 10, adherent fractions were digested with 10 mg/mL collagenase II (Worthington) and 10 mg/mL Nattokinase (Japan BioSciences) for 1 h on a rocker at 37°C. Tissues were further digested a second iteration if not completely detached. Isolated cells were washed and resuspended in FACS buffer 2X and filtered with 40 μm Flowmi® Cell Strainers (Sigma). Cells were then sorted for tdTomato signal using a Sony MA900 cell sorter, collected in RNA lysis buffer, and processed using RNeasy Micro Kit (Qiagen), according to the manufacturer's instructions. RNA was analyzed for quality and quantity using an Agilent Qubit 2.0 bioanalyzer at the Herbert Irving Comprehensive Cancer Center Molecular Pathology Shared Resource at Columbia University (all RIN > 9.5). Samples were sequenced by the Columbia Genome Center.