Fig. 4

CLIP3 controls plasma membrane translocation of GLUT3 in GBM cells. A ECAR was measured by Seahorse analyzer upon transfection of both CD133 and NES siRNA in U87MG and T98G cells. Bar graphs depict glycolysis (measured by the generation of lactate upon glucose addition) and glycolytic capacity (the maximum capacity of lactate generation upon inhibition of oxidative phosphorylation). 2-DG, 2-Deoxy-D-glucose. B mRNA levels of CLIP3, GLUT1, and GLUT3 were analyzed by real-time qRT-PCR upon CLIP3 siRNA treatment in U87MG and T98G cells. C mRNA levels of CLIP3, GLUT1, and GLUT3 were analyzed by real-time qRT-PCR upon transfection of CLIP3 gene in U87MG and T98G cells. D Representative immunofluorescence staining for Spy1 (red), GLUT3 (green), and merged images (with DAPI, blue) on Control, IR (6 Gy), or Spy1 siRNA treatment in U87MG and T98G cells. Scale bars, 10 μm. E Representative immunofluorescence images for Rab11a-OFP (red), GLUT3-GFP (green), and merged images on Control, IR (6 Gy), or CLIP3 siRNA treatment in U87MG and T98G cells. Scale bars, 10 μm. F Confocal live-cell images of U87MG and T98G cells stably expressing GLUT3-GFP after treatment of IR (6 Gy) or CLIP3 siRNA for the indicated times. Arrows indicate GLUT3-GFP-containing vesicular structures migrating toward the plasma membrane. Scale bars, 5 μm. *p < 0.05, **p < 0.01, ***p < 0.001 with unpaired t-test