Fig. 4

PDGFRα and PDGF-D are essential for GBM growth and invasion. A, B sgRNA knockout of PDGFRA or PDGFD in U251 cells was confirmed by Western blot. C, D PDGFRA KO cells and PDGFD KO cells were treated with hypoxia and the expression of HIF1α and others was detected by Western blot. E,F The knockout of PDGFRA or PDGFD inhibits colony growth and cell invasion compared with scrambled sgRNA polyclonal WT cells. KO or WT cell lines were seeded into a 6-well plate (5 × 10³ cells/well) and the reduced growth of the KO cells was determined by colony growth assay (E,F, upper). The KO or WT cell lines were seeded into the upper chamber of a 24-well transwell plate (1 × 10⁵ cells/well) and the cell invasive activity shown in (E,F, lower). G, H PDGFRα and PDGF-D are essential for tumor growth. WT, PDGFRA-KO and PDGFD-KO cells (mixed clones of each) were transduced with GFP-Luciferase, and intracranially transplanted into NSG mice with 5x10⁴cells/mouse for each line. Kaplan-Meier survival curves are shown for the mice. Three independent experiments were performed to confirm the roles of PDGFRα and PDGF-D in tumor cell growth and invasion and tumor formation in mice