Illumina HiSeq 2500 sequencing; ENDOTHELIAL INHIBITION OF THE GLYCOLYTIC ACTIVATOR PFKFB3 INDUCES TUMOR VESSEL NORMALIZATION, IMPAIRS METASTASIS AND IMPROVES CHEMOTHERAPY
ENDOTHELIAL INHIBITION OF THE GLYCOLYTIC ACTIVATOR PFKFB3 INDUCES TUMOR VESSEL NORMALIZATION, IMPAIRS METASTASIS AND IMPROVES CHEMOTHERAPY
Mouse endothelial cells were isolated from perfused healthy liver (for normal endothelial cells, NECs) or liver containing B16-F10 tumors (tumor endothelial cells, TECs). Since the tumor-infested liver consisted for 60-80% of tumor tissue, the EC population isolated from these livers was highly enriched in TECs (therefore referred to as “eTECs”). The purity of cultured endothelial cells was determined by morphological analysis (by phase-contrast microscopy) and by PCR amplification for the EC-enriched surface markers Pecam1 and Eng, and genes enriched in pericytes (Pdgfrb), smooth muscle cells and fibroblasts (A, Pdgfra) and leukocytes (Ptprc). In addition, FACS analysis confirmed that >95% of cultured cells were CD31+ CD102+ CD11b- ECs. Potential contamination of cancer cells was evaluated by FACS using mCherry+ B16 cells (<2% of the total EC population). RNA extraction was performed using the Trizol reagent.  RNA libraries were created using the Illumina TruSeq RNA sample preparation kit V2 according to the manufacturer’s instructions.
Vesalius Research Center, VIB Laboratory of Translational Genetics Department of Oncology, KU Leuven
E-MTAB-4842:ARI007
Experimental Factor: cancer
disease