Fig 1: Active glucose uptake takes place in the eye field territory of mouse embryos.A–C A strong glucose uptake signal was initially detected mostly in the presumptive OVs at E8.0 (arrowheads), and in the midbrain and tail bud region at the 5-6 somite stage (E8.5). Representative micrographs are shown as similar results were obtained from three independent experiments. D–I Glucose tracer uptake was impaired in the presumptive OV territory of Glut1, Rax and Six3 mutant embryos compared with that in their heterozygous littermates. J Quantification of 2-NBDG fluorescence intensity in those embryos. 2-NBDG intensity in the anterior neural plate (ANP) was measured relative to the heart primordium. K–P Bright field images and immunostaining showing that OV morphogenesis in Glut1 and Ldha conditional null embryos was impaired. Rax expression was reduced, while expression of the forebrain marker Lhx5 was not dramatically affected in those mutant embryos. The Delta allele was generated by germline deletion using Ella-Cre mice. Only one flox allele remains upon conditional Cre expression. Q Quantification of Rax expression relative to DAPI signals. R Schematic representation of the glycolysis process during eye morphogenesis. Scale bars, 100 µm (A–I, K–P). Unpaired Student’s t test (two-tailed) was performed (J, Q). ** or *** indicates a p-value is less than 0.01 or 0.001, respectively (J, Q). Data are presented as mean values +/−SEM (J, Q). Source data are provided as a Source Data file. n = 3 biologically independent experiments were performed.
Fig 2: (A) Correlation matrix of AGI compared with immunohistochemistry measurements shown with Spearman’s correlation coefficients. * P < 0.05, ** P < 0.01. Scatterplots of AGI compared with immunohistochemistry measurement of GLUT3 (B), HK2 (C), and MCT1 (D). Lines show linear regression fit to the data. GLUT3 = glucose transporter 3; HK2 = hexokinase 2, LDHA = lactate dehydrogenase A, MCT1 = monocarboxylate transporter 1.
Fig 3: SLC10A1 suppressed aerobic glycolysis in HCC cells. (a) The ECARs of HepG2 and HuH-7 cells with SLC10A1 overexpression were assessed. (b) The OCRs of HepG2 and HuH-7 cells with SLC10A1 overexpression were assessed. (c) The mRNA expression of GLUT1, HK2, LDHA, and PGK1 in HepG2 and HuH-7 cells transfected with pcDNA-3.1-SLC10A1 was determined by qRT-PCR. (d) The protein expression of GLUT1, HK2, LDHA, and PGK1 in HepG2 and HuH-7 cells transfected with pcDNA-SLC10A1 was determined by western blot. ∗∗P < 0.01.
Fig 4: Glycolysis regulates optic vesicle morphogenesis in eye organoids.A Violin plots identify higher expression of the early eye differentiation markers Six3, Rax, Pax6 and Lhx2 on cluster 1. They also express Slc2a1 (Glut1), Slc2a3 (Glut3) and Ldha. Lhx5 is a forebrain marker. B–H Pharmacological inhibition of glucose catabolism and LDH activity by 2-Deoxy-d-Glucose, 2-DG (2 mM) and GNE-140 (20 µM) respectively, shows that formation of RaxGFP+-expressing OVs (arrowheads) is arrested and the expression levels of eye markers such as Rax and Six3 is severely reduced (E, F, H). Insets indicate Rax-GFP expression (D–H). Representative micrographs are shown as similar results were obtained from three independent experiments (C–H). I Quantitative PCR analysis of a set of key eye field transcription factors known to be necessary during the process of eye morphogenesis. Scale bars, 100 µm (C–H). Unpaired Student’s t test (two-tailed) was performed (I). * or *** indicates a p-value is less than 0.05, or 0.001, respectively (I). Data are presented as mean values +/−SEM (I). Source data are provided as a Source Data file. n = 3 biologically independent experiments were performed.
Fig 5: Schematic overview of metabolic pathway in IDH wild-type and mutant gliomas. Similar to other malignant tumors, IDH wild-type glioma is characterized by a high level of aerobic glycolysis (Warburg effect). Hypoxia induces upregulation of HIF1α, leading to overexpression of glycolysis-related proteins/enzymes, namely, GLUT, HK2, MCT, and LDHA, thereby accelerating glycolysis. On the other hand, HIF1α response to hypoxia is blunted in IDH mutant glioma, shifting the metabolism to oxidative phosphorylation. In this study, the extent of aerobic glycolysis was estimated by pH- and oxygen-sensitive imaging combined with perfusion imaging. The proteins/enzymes investigated in our study are labeled in black. Notably, MTRasym at 3.0 ppm increases not only in accordance with decrease in pH but also with increase in glutamine related to tumor activity. 2HG = 2-hydroxyglutarate, aKG = alpha-ketoglutarate, acetyl-CoA = acetyl coenzyme A, G6P = glucose-6-phosphate, GDH = glutamate dehydrogenase, GLS = glutaminase, GLUT = glucose transporter, HK2 = hexokinase 2, LAT = L-amino acid transporter, LDHA = lactic dehydrogenase A, MCT = monocarboxylate transporter, mTOR = mammalian target of rapamycin, PDH = pyruvate dehydrogenase, PDK1 = pyruvate dehydrogenase kinase 1, ROS = reactive oxygen species, TA = transaminase, TCA cycle = tricarboxylic acid cycle.
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