Fig. 4
6-Phosphogluconate dehydrogenase (6PGD) Y481 phosphorylation promotes DNA synthesis and glioma progression. a U87/EGFRvIII cells stably expressing luciferase were depleted of endogenous 6PGD and reconstituted with the expression of r6PGD WT or Y481F. The expression of 6PGD was examined using immunoblotting assays. b–e NADP+/NADPH ratio (b), relative reactive oxygen species (ROS) levels (c), cellular ribulose-5-phosphate (Ru-5-P) level (d), and BrdU incorporation ratio (e) were measured in these cells generated in a. f The flux through the pentose phosphate pathway (PPP) was determined using d-glucose-1,2-13C2 in these cells generated in a. g Cellular ATP levels were determined in these cells generated in a. h, i Cell proliferation (h) and colony formation (i) were determined in these cells generated in a. j, k These cells generated in a (2 × 105 per mouse) were intracranially injected into randomized athymic nude mice (five mice per group). Bioluminescence imaging of tumor growth were carried out. Real time images were presented (j, left panel) and the intensities of luciferase were quantified (j, right panel). After 11 days, tumor growth was examined. Hematoxylin and eosin (H&E)-stained coronal brain sections show representative tumor xenografts. Scale bar, 100 μm (k, left panel). Representative images of tumor boundaries were presented with ×200 magnification. Tumor volumes were measured using length (a) and width (b) and calculated using the equation: V = ab2/2 (k, right panel). Data represent the mean ± SD of luciferase intensity of five mice per group. Student’s t-test (unpaired, two tailed), **p < 0.01. b–i Data represent the mean ± SD of three independent experiments. Student’s t-test (unpaired, two tailed), **p < 0.01; n.s. not significant. Source data are provided as a Source Data file
