title: GSM8518539: BMDM 3 treated with vehicle control; Mus musculus; RNA-Seq
library_name: GSM8518539
library_construction_protocol: Bone marrow was flushed from the hind tibia and femurs of 8-week old female Balb/CJ mice using the protocol detailed in (doi: 10.3791/53936). After which bone marrow was filtered with at 40-micron filter and pelleted via centrifugation. Pellets were treated with 2 mLs ACK (150 mM Ammonium Chloride, 1 mM Potassium Bicarbonate, 0.1 mM EDTA, pH 7.4) to lyse red blood cells that was quenched with the addition of 10 mLs of HBSS. Cells were washed 2x with HBSS (400 mg/L KCl, 60 mg/L KH2PO4, 350 mg/L NaHCO3, 8000 mg/L NaCl, 48 ng/L Na2HPO4, 1,100 mg/L dextrose anhydrase) then resuspended in 10 mLs growth media (RMPI + 10% FBS + 1% pen/strep) and plated (~1.125-2 x 10^6 cells per petri dish). To differentiate cells into macrophages, they were treated with 25 ng/mL M-CSF (Shenandoah, 200-08) upon plating and again 2 days later. After 5 days, BMDM were treated with 2.5 uM bilirubin or DMSO control. After 48 hours treatment with bilirubin, mRNA was isolated from BMDM using TriZOL extraction as recommended by the manufacturer (FisherScientific, 15596-018). Poly A selected total RNA library was prepared after mRNA extraction
identifiers.namespace: GEO
identifiers.id: GSM8518539_r1