RNAseq of tumor endothelial cells isolated from CT2A glioma under control or LGK974 treatment

The Wnt signaling pathway regulates blood-brain barrier (BBB) formation and function in both development and pathogenesis of diseases, and is essential for CNS angiogenesis and BBB formation in both embryonic and postnatal development. However, how Wnt signaling affects brain tumor endothelial cells is largely unknown. We isolated endothelial cells from tumors in control or LGK974 treated mice, and analyzed the role of Wnt signaling inhibition on tumor endothelial cells. We injected GFP-labeled CT2A glioma cells intracranially to C57BL/6 mice of 6 weeks old. 21 days after tumor induction, mice were administrated with saline or 2.5mg/kg LGK974 by oral gavage for 4 consecutive days. 25 days after tumor injection, tumor were dissected under a stereological microscope according to GFP expression. Magnetic-activated cell sorting (MACS) was used for isolating endothelial cells, based on expression of CD31 and CD45 (CD45-/CD31+), from tumors in control or LGK974 treated mice. Isolated endothelial cells were further prepared for RNAseq analysis.