GSM2653227: CSF1R-49_L5; Mus musculus; RNA-Seq
Tumors were excised and single cell suspensions obtained by mechanical processing and enzymatical digestion (DNase 0.01%, Dispase 1mg/ml, Collagenase IV 1mg/ml) 16 hrs after treatment. TAM (CD11b+F4/80hiLy6ClowLy6Gneg), monocytic MDSC (CD11b+F4/80-/lowLy6ChiLy6Gneg) were sorted using flow cytometry.  RNA was extracted from multiple FACS-sorted cells or from whole tumor. 1 ng of amplified cDNA was subjected to Nextera XT library preparation (Illumina) according to manufacturer’s instructions. Sequencing libraries were quantified using the Kapa Library Quantification kit (Kapa Biosystems) and quality controlled by capillary electrophoresis on a Bioanalyzer using High Sensitivity chips (Agilent Technologies). Libraries were sequenced on a HiSeq2500 sequencer (Illumina) for 2 x 50 cycles using version 4 cluster generation kits and version 4 sequencing reagents (Illumina).
GEO Accession
GSM2653227