Journal: BMC Pulmonary Medicine

Article Title: FUT3 facilitates glucose metabolism of lung adenocarcinoma via activation of NF-κB pathway

doi: 10.1186/s12890-023-02688-x

Figure Lengend Snippet: FUT3 participated in glucose metabolism in LUAD cells. (a) GLUT1 and LDHA immunofluorescence was performed to present the alteration of glucose metabolism in H1975 and SPCA-1 cells after FUT3 knockdown. (b-c) The enzyme activity of G6PDH and LDH was measured in LUAD cells. (* P ＜0.05, ** P < 0.01, **** P < 0.0001)

Article Snippet: Cells were seeded in a 24-well plate at a density of 5.0 × 10 3 cells/well, fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.5% Triton X-100 for 5 min. After blocked with 5% fetal bovine serum for 1 h, cells were incubated with primary antibodies of GLUT1 (Cat.# ab115730, 1:200, abcam) and LDHA (Cat.# ab47010, 1:1000, abcam) overnight at 4 °C.

Techniques: Immunofluorescence, Activity Assay

Journal: PLoS Biology

Article Title: Structural myelin defects are associated with low axonal ATP levels but rapid recovery from energy deprivation in a mouse model of spastic paraplegia

doi: 10.1371/journal.pbio.3000943

Figure Lengend Snippet: (A) Immunoblot analysis of myelin purified from the brains of wild-type (control [Ctrl]) and Plp null/y mice probed for glucose transporter 1 (GLUT1) and monocarboxylate transporter 1 (MCT1). β-actin (ACTB) was detected as a loading control; PLP/DM20 was detected as genotype control. Blot represents N = 4 individual mice for each genotype. (B and C) Genotype-dependent quantification of the immunoblots for GLUT1 (B) and MCT1 (C). The abundance of both proteins is significantly increased in myelin purified from Plp null/y brains ( N = 4 mice for both genotypes). (D and E) Quantitative RT-PCR analysis of the abundance of Glut1 (D) and Mct1 (E) mRNAs in the corpus callosum dissected from wild-type (Ctrl) and Plp null/y mice ( N = 4 mice for both genotypes). In (B–E): ** p < 0.01, *** p < 0.001; Student t test. (F–I) Cryo-immuno electron microscopy to assess the localization of GLUT1 (F and G) and MCT1 (H and I) protein in the myelin of optic nerves dissected from wild-type (F and H) and Plp null/y (G and I) mice. White arrows point at gold particles indicating localization of the respective proteins. The scale bars in (F–I) correspond to 200 nm. White boxes are 2.6× zoomed-in magnification of the corresponding box highlighting the gold particles. Original, uncropped Western blots are available in . Data underlying this figure can be found in .

Article Snippet: Primary antibodies were specific for GLUT1 (1:1,000) [ ], MCT1 (1:1,000) [ ], β-actin (ACTB; 1:1,000, MAB1501, Merck Millipore), and PLP/DM20 (1:5,000, A431) [ ].

Techniques: Western Blot, Purification, Control, Quantitative RT-PCR, Immuno-Electron Microscopy

Journal:

Article Title: GLUT1 Expression Is Increased in Hepatocellular Carcinoma and Promotes Tumorigenesis

doi: 10.2353/ajpath.2009.080596

Figure Lengend Snippet: Glut1 Immunoreactivity (IR) in HCC Tissue of 152 Patients in Relation to Clinicopathological Characteristics and Proliferation Rate

Article Snippet: Protein extraction and Western blotting were performed as described elsewhere, 27 applying the following primary antibodies: anti-Glut1 (1:600; Lab Vision, Wedel, Germany), anti-HIF-1 α (1:500; Novus Biologicals, Littleton, CO), and anti-β-actin (1:20,000, Sigma).

Techniques:

Journal:

Article Title: GLUT1 Expression Is Increased in Hepatocellular Carcinoma and Promotes Tumorigenesis

doi: 10.2353/ajpath.2009.080596

Figure Lengend Snippet: GLUT1 expression in HCC. GLUT1 mRNA (A) and protein (B) expression in PHHs and three different HCC cell lines (HepG2, PLC, and Hep3B) analyzed by quantitative real-time PCR (A) and Western blotting (B). Data are given as mean ± SEM (*P < 0.05). C: GLUT1 mRNA expression in 22 human HCC samples in relation to matching nontumorous liver tissue samples. D: Glut1 immunohistochemical staining of human HCC tissue and adjacent nontumorous liver tissue (nt) and higher magnification of the same HCC tissue sample. E: Proliferation rate (analyzed by immunohistochemical staining applying anti-Ki-67 antibodies) in HCC-tissues with positive or negative immunoreactivity to Glut1 applying TMA technology. Original magnifications: ×40 (left); ×400 (right).

Article Snippet: Protein extraction and Western blotting were performed as described elsewhere, 27 applying the following primary antibodies: anti-Glut1 (1:600; Lab Vision, Wedel, Germany), anti-HIF-1 α (1:500; Novus Biologicals, Littleton, CO), and anti-β-actin (1:20,000, Sigma).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining, Staining

Journal:

Article Title: GLUT1 Expression Is Increased in Hepatocellular Carcinoma and Promotes Tumorigenesis

doi: 10.2353/ajpath.2009.080596

Figure Lengend Snippet: Regulation of GLUT1 expression in HCC under aerobic and anaerobic conditions. A: HIF-1α expression in three different HCC cell lines (HepG2, PLC, and Hep3B) with or without pharmacological induction of hypoxia by DP (100 μmol/L) analyzed by Western blotting. B: Luciferase activity in Hep3B cells transiently transfected with a reporter gene driven by six hypoxia-responsive elements (HREs) in relation to cells transfected with control plasmid (pcDNA3). DP (2,2′-dipyridy, 100 μmol/L), YC-1 (100 μmol/L); ECH (echinomycin, 10 nmol/L). * and #, P < 0.05 compared with control and DMSO without DP or with DP, respectively. C: GLUT1 mRNA expression in Hep3B cells with or without pharmacological induction of hypoxia or HIF-1α inhibition analyzed by quantitative real-time PCR. * and #, P < 0.05 compared with control and DMSO without DP or with DP, respectively. D: Analysis of GLUT1 mRNA expression in HCC cells cultured under normoxic conditions or 1% oxygen for 16 hours. *P < 0.05 compared with control. E: Hep3B cells were exposed to normoxic or hypoxic conditions, actinomycin D (Act.D, 7.5 μg/ml) was added, and incubation was continued for 3, 6, and 12 hours. GLUT1 mRNA was analyzed by qPCR. F: Analysis of GLUT1 mRNA expression in Hep3B cell transiently transfected with two different MAZ siRNAs (siRNA1 and siRNA2), and cells transfected with control siRNA and nontransfected HCC cells (ctrl.). *P < 0.05 compared with control. All experiments have been performed at least three times. Data are given as mean ± SEM.

Article Snippet: Protein extraction and Western blotting were performed as described elsewhere, 27 applying the following primary antibodies: anti-Glut1 (1:600; Lab Vision, Wedel, Germany), anti-HIF-1 α (1:500; Novus Biologicals, Littleton, CO), and anti-β-actin (1:20,000, Sigma).

Techniques: Expressing, Western Blot, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Inhibition, Real-time Polymerase Chain Reaction, Cell Culture, Incubation

Journal:

Article Title: GLUT1 Expression Is Increased in Hepatocellular Carcinoma and Promotes Tumorigenesis

doi: 10.2353/ajpath.2009.080596

Figure Lengend Snippet: Effect of GLUT1 inhibition on migration and proliferation of HCC cells. Analysis of GLUT1 (A) mRNA and (B) protein expression in Hep3B cells transiently transfected with two different GLUT1 siRNAs (siRNA1 and siRNA2), and cells transfected with control siRNA and nontransfected HCC cells (ctrl.). C: The migratory potential of these cells was assessed by Boyden chamber assays. D: For analysis of proliferation, cells were trypsinized and counted at different time points. Cell number at day 4 is set at 1. E: Growth in a three-dimensional cell culture system was assessed by analysis of the spheroid volume at day 14. All experiments have been performed at least three times. Data are given as mean ± SEM (*P < 0.05 compared with controls).

Article Snippet: Protein extraction and Western blotting were performed as described elsewhere, 27 applying the following primary antibodies: anti-Glut1 (1:600; Lab Vision, Wedel, Germany), anti-HIF-1 α (1:500; Novus Biologicals, Littleton, CO), and anti-β-actin (1:20,000, Sigma).

Techniques: Inhibition, Migration, Expressing, Transfection, Cell Culture

Journal:

Article Title: GLUT1 Expression Is Increased in Hepatocellular Carcinoma and Promotes Tumorigenesis

doi: 10.2353/ajpath.2009.080596

Figure Lengend Snippet: Effect of GLUT1 on rates of glucose uptake and glycolysis in HCC. Analysis of lactate secretion into the supernatant of PHHs and three different HCC cell lines (HepG2, PLC, and Hep3B) (A) and Hep3B cells transiently transfected with two different GLUT1 siRNAs (siRNA1 and siRNA2) (B), cells transfected with control siRNA and nontransfected HCC cells (ctrl.). C: Glucose uptake is reported as glucose utilization per cell within 24 hours. To that end, glucose concentration was measured in the supernatant and cells were counted 24 hours after seedings. All experiments have been performed at least three times. Data are given as mean ± SEM (*P < 0.05 compared with controls).

Article Snippet: Protein extraction and Western blotting were performed as described elsewhere, 27 applying the following primary antibodies: anti-Glut1 (1:600; Lab Vision, Wedel, Germany), anti-HIF-1 α (1:500; Novus Biologicals, Littleton, CO), and anti-β-actin (1:20,000, Sigma).

Techniques: Transfection, Concentration Assay

Journal: Cancer Medicine

Article Title: Identification and Experimental Validation of Prognostic miRNA Signature and Ferroptosis‐Related Key Genes in Cervical Squamous Cell Carcinoma

doi: 10.1002/cam4.70415

Figure Lengend Snippet: Target genes for seven miRNAs and consensus genes and key genes in CESC. (A) miR‐505‐5p, (B) miR‐425‐5p, (C) miR‐331‐3p, (D) miR‐502‐3P, (E) miR‐629‐3p, (F) miR‐100‐3p, (G) miR‐301a‐5p. (H) The overlap between 86 DEGs and 176 target genes. (I) The association of SLC2A1 with patients' prognosis of cervical cancer. (J) The association of TXNIP with patients' prognosis of cervical cancer. (K) The association of ANO6 with patients' prognosis of cervical cancer. (L) Network shows the intricate interplay between miRNAs and their target genes. Red denotes upregulated; green denotes downregulated.

Article Snippet: Thereafter, the membranes were incubated with primary antibodies directed against SLC2A1 (1:500, ZENBIO, 380464), TXNIP (1:500, Abcam, ab188865), and GAPDH (1:10,000, Abcam, ab8245) overnight.

Techniques:

Journal: Cancer Medicine

Article Title: Identification and Experimental Validation of Prognostic miRNA Signature and Ferroptosis‐Related Key Genes in Cervical Squamous Cell Carcinoma

doi: 10.1002/cam4.70415

Figure Lengend Snippet: 12 consensus genes.

Article Snippet: Thereafter, the membranes were incubated with primary antibodies directed against SLC2A1 (1:500, ZENBIO, 380464), TXNIP (1:500, Abcam, ab188865), and GAPDH (1:10,000, Abcam, ab8245) overnight.

Techniques:

Journal: Cancer Medicine

Article Title: Identification and Experimental Validation of Prognostic miRNA Signature and Ferroptosis‐Related Key Genes in Cervical Squamous Cell Carcinoma

doi: 10.1002/cam4.70415

Figure Lengend Snippet: Biological functions of differentially expressed genes and key genes. (A) Bar plots showing GO analysis for BP. (B) Bar plots showing GO analysis for MF. (C) Bar plots showing GO analysis for CC. (D) Circos plots of KEGG analysis. (E) Box plots of expression levels of key genes. * p < 0.05, ** p < 0.01, **** p < 0.001. (F) The single‐gene GSEA analysis of SLC2A1. (G) The single‐gene GSEA analysis of TXNIP.

Article Snippet: Thereafter, the membranes were incubated with primary antibodies directed against SLC2A1 (1:500, ZENBIO, 380464), TXNIP (1:500, Abcam, ab188865), and GAPDH (1:10,000, Abcam, ab8245) overnight.

Techniques: Expressing

Journal: Cancer Medicine

Article Title: Identification and Experimental Validation of Prognostic miRNA Signature and Ferroptosis‐Related Key Genes in Cervical Squamous Cell Carcinoma

doi: 10.1002/cam4.70415

Figure Lengend Snippet: Drug sensitivity and experimental verification of miRNA signature and key genes. (A) Scatter diagram of the relationship between consensus genes and drug sensitivity. (B) Network diagram of miRNA and target gene and drug correlation. (C) Differential expression of miR‐301a‐5p between cervical squamous cell carcinoma tumor and paracancerous tissues. (D) Differential expression of miR‐505‐5p between cervical squamous cell carcinoma tumor and paracancerous tissues. (E) SLC2A1 expression in cervical cancer and paracancerous tissues. (F) TXNIP expression in cervical cancer and paracancerous tissues. (G) The Stripe plots of Western Blotting analysis. * p < 0.05, ** p < 0.01, **** p < 0.001.

Article Snippet: Thereafter, the membranes were incubated with primary antibodies directed against SLC2A1 (1:500, ZENBIO, 380464), TXNIP (1:500, Abcam, ab188865), and GAPDH (1:10,000, Abcam, ab8245) overnight.

Techniques: Quantitative Proteomics, Expressing, Western Blot

Journal: Frontiers in Neuroscience

Article Title: Diffusion Tensor Imaging and Chemical Exchange Saturation Transfer MRI Evaluation on the Long-Term Effects of Pulsed Focused Ultrasound and Microbubbles Blood Brain Barrier Opening in the Rat

doi: 10.3389/fnins.2020.00908

Figure Lengend Snippet: Representative immunohistochemistry (IHC) staining of the endothelial glucose transporter 1 (GLUT1), neuronal glucose transporter 3 (GLUT3), and neuronal nuclei (NeuN) of a Group 2 rat brain that was obtained at week 7 after six pFUS+MB treatments. Dilated vessels were found in the treated hemisphere (asterisk). The IHC images were acquired and quantified from the (A,B) cortex and (C,D) hippocampus. * p < 0.05, ** p < 0.01 vs. contralateral untreated region, paired t -test ( n = 3/stain/group).

Article Snippet: The following primary antibodies were used: glucose transporter 1 (GLUT1) (Invitrogen MA5-11315, Waltham, MA) at 1/200; glucose transporter 3 (GLUT3) (Abcam ab41525, Cambridge, United Kingdom) at 1/1500; hexaribonucleotide binding protein-3 (NeuN) (Cat. mab377, Millipore) at 1/1000.

Techniques: Immunohistochemistry, Staining