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  • 96
    Synaptic Systems guinea pig anti vglut1
    Guinea Pig Anti Vglut1, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 96/100, based on 252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore glut1
    Glut1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 459 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam slc2a1
    Slc2a1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti glut1
    <t>GLUT1</t> expression vector construction
    Anti Glut1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 379 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher glut1
    Kaplan-Meier estimates of survival functions for DFS in the low and high groups for the SUVmean, SUVmax, MTV, TLG, <t>Glut1,</t> pStat1, and pStat3 in the 140 NSCLC patients P-values were determined by the log-rank test.
    Glut1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 389 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alpha Diagnostics glut1
    ( a–f ) Role of <t>GLUT1</t> in VEGF-induced MC FN production. ( a ) Primary culture mouse MCs with the C57BL6 background were studied in culture with and without VEGF-A 165 peptide (2 ng/ml) for 48 h. VEGF-A 165 stimulated a two-fold increase in MC GLUT1
    Glut1, supplied by Alpha Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 183 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti glucose transporter glut1 antibody
    Effects of co-culture time and 3T3-Ll cell density on mRNA levels of Glutl and Glut4 in L6 cells. L6 cells were incubated with 3T3-Ll adipocytes at various times at 5 × 10 3 density and mRNA levels of (A) <t>Glut1</t> and (B) Glut4 measured. L6 cells were incubated with 3T3-Ll adipocytes for 6 h at indicated densities and the mRNA levels of Glutl . P values by one-way ANOVA followed by post-hoc Tukey HSO test are shown. ANOVA P
    Anti Glucose Transporter Glut1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti glut1
    Representative image of immunostaining corresponding to the glucose transporter protein <t>SLC2A1</t> <t>(GLUT1)</t> in 30 week-old Ins2 AKITA and WT cerebral cortex. Mural cells (ANPEP, green), glucose transporter 1 (GLUT1, red), endothelium (PECAM1, cyan). n = 2, scale bar 50 µm.
    Anti Glut1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 280 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Proteintech glut1
    The image analysis results showed that the expression of <t>GLUT1</t> on membrane was detected in cells transfected with cab39. Compared with un-transfected cells, the expression rates of GLUT1 in H460 cells (A) and LK2 cells (B) were 45.5% and 67.1%, respectively. Efficiency was measured by Western blotting. (C) The results of flow cytometry showed that the average density of glucose in H460 cells transfected with cab39 increased by 3.3 times ( p
    Glut1, supplied by Proteintech, used in various techniques. Bioz Stars score: 90/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti glucose transporter glut1 antibody epr3915
    The image analysis results showed that the expression of <t>GLUT1</t> on membrane was detected in cells transfected with cab39. Compared with un-transfected cells, the expression rates of GLUT1 in H460 cells (A) and LK2 cells (B) were 45.5% and 67.1%, respectively. Efficiency was measured by Western blotting. (C) The results of flow cytometry showed that the average density of glucose in H460 cells transfected with cab39 increased by 3.3 times ( p
    Anti Glucose Transporter Glut1 Antibody Epr3915, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Merck KGaA glut1
    Insulin does not stimulate GLUT or IR protein expression in villous explants from term placentas of women with obesity. Villous explants from term placentas were treated with and without insulin, and (A) MVM <t>GLUT1,</t> (B) BM GLUT1 (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression was measured by western blot (n = 6 per group). Data are shown as mean ± SEM. Significance was evaluated with a paired t test.
    Glut1, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rabbit anti glut1
    <t>Glut1</t> expression increases GSK-3 phosphorylation, Mcl-1 level, and survival in primary hematopoietic cells. (A) Bone marrow cells were cultured in IL-3, infected with retrovirus to express Glut1 and truncated hNGFR as a marker on day 4, and stained by immunofluorescence for phospho-GSK-3 (red), hNGFR (green), and DAPI on day 7. (B to E) Glut1 protein levels were analyzed by immunoblotting (B), glucose uptake was measured (C), and phospho-GSK-3 levels were determined by immunoblotting (D) and quantified (E) in purified T lymphocytes from nontransgenic (Non) and Glut1-transgenic (Glut1) littermates freshly after purification or after 1 day of culture in the absence of stimulation (neglect). (F) Mcl-1 levels were determined by immunoblotting of freshly isolated, neglected, or GSK-3 inhibitor (10 μM SB216763 [SB])-treated nontransgenic T cells. Ratios are shown. (G) Mcl-1/actin ratio quantitated from nontransgenic and Glut1-transgenic T cells after isolation (fresh), neglect, or culture in GSK-3 inhibitor (SB). (H) Purified T cells from nontransgenic and Glut1-transgenic littermate mice were cultured without stimulation (neglect), and cell survival determined after 2 days. Values shown are the means from triplicate samples; error bars indicate standard deviations.
    Rabbit Anti Glut1, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 184 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc glut1
    HOXA9 represses the expression of HIF-1α and its downstream glycolytic genes by directly binding to the promoter regions. a , b The mRNA or protein expression of HOXA9, HIF-1α, HK2, <t>GLUT1,</t> and PDK1 was detected by qRT-PCR or western blot, respectively, in A431 cells after depletion of HOXA9 by two siRNAs or overexpression of HOXA9. The qRT-PCR data were normalized to GAPDH gene expression. In western blots, GAPDH was used as a loading control. c Known or predicted binding sites of HOXA9 (diamond) or HRE elements for HIF-1α (delta) in the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 ). d Electrophoretic mobility shift assay (EMSA) was used to detect the direct association of purified HOXA9 protein with its binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 . The black arrows indicate the binding complex of the HOXA9 protein with the probe containing the known or predicted binding sites whereas the white arrows indicate the supershift generated by the association of the anti-HOXA9 antibody with the above complexes. Also, the specificity of the direct associations was supported by the loss of binding activities using mutated probes. e The binding enrichment of HOXA9 at the above binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 was detected by ChIP-PCR after knockdown of HOXA9. f The binding enrichment of HIF-1α at the above binding sites on the promoter regions was detected after overexpression of HOXA9. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P
    Glut1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher glut 1 rabbit polyclonal antibody
    HOXA9 represses the expression of HIF-1α and its downstream glycolytic genes by directly binding to the promoter regions. a , b The mRNA or protein expression of HOXA9, HIF-1α, HK2, <t>GLUT1,</t> and PDK1 was detected by qRT-PCR or western blot, respectively, in A431 cells after depletion of HOXA9 by two siRNAs or overexpression of HOXA9. The qRT-PCR data were normalized to GAPDH gene expression. In western blots, GAPDH was used as a loading control. c Known or predicted binding sites of HOXA9 (diamond) or HRE elements for HIF-1α (delta) in the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 ). d Electrophoretic mobility shift assay (EMSA) was used to detect the direct association of purified HOXA9 protein with its binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 . The black arrows indicate the binding complex of the HOXA9 protein with the probe containing the known or predicted binding sites whereas the white arrows indicate the supershift generated by the association of the anti-HOXA9 antibody with the above complexes. Also, the specificity of the direct associations was supported by the loss of binding activities using mutated probes. e The binding enrichment of HOXA9 at the above binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 was detected by ChIP-PCR after knockdown of HOXA9. f The binding enrichment of HIF-1α at the above binding sites on the promoter regions was detected after overexpression of HOXA9. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P
    Glut 1 Rabbit Polyclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals glut1 antibody
    Graphical representation of PrP C -mediated modulation of glucose homeostasis through iron. (1 2) PrP C facilitates cellular uptake of Tf-Fe 3+ and non-Tf-bound iron (Fe 3+ . (3) , resulting in the downregulation of GLUT2 in pancreatic β-cells and hepatocytes (black), GLUT 3 in neuronal cells (red), and <t>GLUT1</t> at the blood-retinal barrier (blue). (4) Reduced uptake of glucose downregulates insulin, resulting in hyperglycemia. (5) Down-regulation or deletion of PrP C and glucose transporters. (6) Increased uptake of glucose stimulates insulin secretion with resultant hypoglycemia.
    Glut1 Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human glut1 antibody
    <t>CD4+Glut1+</t> T cell percentage inversely correlates with CD4+ T cell counts (A,B) , CD4+ T cell percentage (C,D) , and CD4/CD8 ratio (E,F) ). cART, combination antiretroviral therapy.
    Human Glut1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam glucose transporter 1 glut1
    Synovial tissue (ST) angiogenesis, oxidative stress and cellular bioenergetics. To support the concept that oxidative stress, angiogenesis and energy metabolism are interconnected processes that co-exist during the inflammation milieu, double-immunofluorescence staining was performed. ST slides were co-incubated with primary mouse antibody against human 4-hydroxy-2-nonenal (4-HNE) and with primary rabbit antibodies against angiogenic factors (vascular endothelial growth factor [VEGF], angiopoietin 2 [Ang2], tyrosine kinase receptor [Tie2]), glycolytic proteins (glyceraldehyde 3-phosphate dehydrogenase [GAPDH], pyruvate kinase isozyme 2 [PKM2], glucose <t>transporter</t> 1 <t>[GLUT1])</t> and a mitochondrial marker (adenosine triphosphate synthase subunit β [ATP5B]). Representative merged immunofluorescence images demonstrate examples of co-localisation ( yellow ) of 4-HNE with VEGF, Ang2, Tie2, GAPDH, PKM2, GLUT1 and ATP5B. Cells stained green are positive for 4-HNE only; cells stained red are positive only for VEGF, Ang2, Tie2, GAPDH, PKM2, GLUT1 and ATP5B. Arrows indicate examples of co-localisation. Magnification of photomicrographs × 20, insets : Figure S4
    Glucose Transporter 1 Glut1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    GLUT1 expression vector construction

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Curcumin inhibits lung cancer invasion and metastasis by attenuating GLUT1/MT1-MMP/MMP2 pathway

    doi:

    Figure Lengend Snippet: GLUT1 expression vector construction

    Article Snippet: After washing, specific antibodies against GLUT1 (Abcam), MT1-MMP (Daiichi Fine Chemical), MMP2 (Abcam) were applied to incubate the membranes at 4°C for 12 hours to detect corresponding proteins.

    Techniques: Expressing, Plasmid Preparation

    Construction and identification of pcDNA3.1-GLUT1. The left part of this figure demonstrates the PCR analysis of GLUT1 full length cDNA fragment. Lane M indicates DNA marker. The right part of this figure shows the pcDNA3.1-GLUT1 vector construction diagram.

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Curcumin inhibits lung cancer invasion and metastasis by attenuating GLUT1/MT1-MMP/MMP2 pathway

    doi:

    Figure Lengend Snippet: Construction and identification of pcDNA3.1-GLUT1. The left part of this figure demonstrates the PCR analysis of GLUT1 full length cDNA fragment. Lane M indicates DNA marker. The right part of this figure shows the pcDNA3.1-GLUT1 vector construction diagram.

    Article Snippet: After washing, specific antibodies against GLUT1 (Abcam), MT1-MMP (Daiichi Fine Chemical), MMP2 (Abcam) were applied to incubate the membranes at 4°C for 12 hours to detect corresponding proteins.

    Techniques: Polymerase Chain Reaction, Marker, Plasmid Preparation

    Effects of curcumin on metastasis of untransfected, empty pcDNA3.1 vector transfected and pcDNA3.1-GLUT1 transfected A549 cells original primary tumor in vivo

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Curcumin inhibits lung cancer invasion and metastasis by attenuating GLUT1/MT1-MMP/MMP2 pathway

    doi:

    Figure Lengend Snippet: Effects of curcumin on metastasis of untransfected, empty pcDNA3.1 vector transfected and pcDNA3.1-GLUT1 transfected A549 cells original primary tumor in vivo

    Article Snippet: After washing, specific antibodies against GLUT1 (Abcam), MT1-MMP (Daiichi Fine Chemical), MMP2 (Abcam) were applied to incubate the membranes at 4°C for 12 hours to detect corresponding proteins.

    Techniques: Plasmid Preparation, Transfection, In Vivo

    Effects of pcDNA3.1-GLUT1 transfection on expressions of GLUT1, MT1-MMP and MMP2 and cell invasion in A549 cells. A. The left part showed the captured image of transwell assay of untrasfected, empty vector transfected and pcDNA3.1-GLUT1 transfected A549

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Curcumin inhibits lung cancer invasion and metastasis by attenuating GLUT1/MT1-MMP/MMP2 pathway

    doi:

    Figure Lengend Snippet: Effects of pcDNA3.1-GLUT1 transfection on expressions of GLUT1, MT1-MMP and MMP2 and cell invasion in A549 cells. A. The left part showed the captured image of transwell assay of untrasfected, empty vector transfected and pcDNA3.1-GLUT1 transfected A549

    Article Snippet: After washing, specific antibodies against GLUT1 (Abcam), MT1-MMP (Daiichi Fine Chemical), MMP2 (Abcam) were applied to incubate the membranes at 4°C for 12 hours to detect corresponding proteins.

    Techniques: Transfection, Transwell Assay, Plasmid Preparation

    Effects of curcumin incubation on expressions of GLUT1, MT1-MMP and MMP2 and cell invasion in A549 cells. (A) The upper part demonstrates the results acquired from transwell assay when A549 cells were incubated with curcucmin of concentrations at 0, 15

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Curcumin inhibits lung cancer invasion and metastasis by attenuating GLUT1/MT1-MMP/MMP2 pathway

    doi:

    Figure Lengend Snippet: Effects of curcumin incubation on expressions of GLUT1, MT1-MMP and MMP2 and cell invasion in A549 cells. (A) The upper part demonstrates the results acquired from transwell assay when A549 cells were incubated with curcucmin of concentrations at 0, 15

    Article Snippet: After washing, specific antibodies against GLUT1 (Abcam), MT1-MMP (Daiichi Fine Chemical), MMP2 (Abcam) were applied to incubate the membranes at 4°C for 12 hours to detect corresponding proteins.

    Techniques: Incubation, Transwell Assay

    Curcumin’s proliferation inhibition effect on A549 cells. Columns in this figure indicated the cell inhibition rate of untransfected, empty vector transfected and pcDNA3.1-GLUT1 transfected A549 cells when incubated with curcumin at serial concentrations

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Curcumin inhibits lung cancer invasion and metastasis by attenuating GLUT1/MT1-MMP/MMP2 pathway

    doi:

    Figure Lengend Snippet: Curcumin’s proliferation inhibition effect on A549 cells. Columns in this figure indicated the cell inhibition rate of untransfected, empty vector transfected and pcDNA3.1-GLUT1 transfected A549 cells when incubated with curcumin at serial concentrations

    Article Snippet: After washing, specific antibodies against GLUT1 (Abcam), MT1-MMP (Daiichi Fine Chemical), MMP2 (Abcam) were applied to incubate the membranes at 4°C for 12 hours to detect corresponding proteins.

    Techniques: Inhibition, Plasmid Preparation, Transfection, Incubation

    Kaplan-Meier estimates of survival functions for DFS in the low and high groups for the SUVmean, SUVmax, MTV, TLG, Glut1, pStat1, and pStat3 in the 140 NSCLC patients P-values were determined by the log-rank test.

    Journal: Oncotarget

    Article Title: The assessment of correlation and prognosis among 18F-FDG uptake parameters, Glut1, pStat1 and pStat3 in surgically resected non-small cell lung cancer patients

    doi: 10.18632/oncotarget.25865

    Figure Lengend Snippet: Kaplan-Meier estimates of survival functions for DFS in the low and high groups for the SUVmean, SUVmax, MTV, TLG, Glut1, pStat1, and pStat3 in the 140 NSCLC patients P-values were determined by the log-rank test.

    Article Snippet: For Glut1, the BenchMark XT was used.

    Techniques:

    Example of NSCLC (adenocarcinoma; pT2N0M0 stage IIA) in an 81-yr-old man High 18 F-FDG uptake is observed in the right upper lung field. (A) MIP image. (B) Axial PET image. SUVmean: 5.39, SUVmax: 8.30, MTV: 110.3, and TLG: 595.31. (C) The Glut1 score was 210. (D) The pStat1 expression score was 1. (E) The pStat3 expression score was 30.

    Journal: Oncotarget

    Article Title: The assessment of correlation and prognosis among 18F-FDG uptake parameters, Glut1, pStat1 and pStat3 in surgically resected non-small cell lung cancer patients

    doi: 10.18632/oncotarget.25865

    Figure Lengend Snippet: Example of NSCLC (adenocarcinoma; pT2N0M0 stage IIA) in an 81-yr-old man High 18 F-FDG uptake is observed in the right upper lung field. (A) MIP image. (B) Axial PET image. SUVmean: 5.39, SUVmax: 8.30, MTV: 110.3, and TLG: 595.31. (C) The Glut1 score was 210. (D) The pStat1 expression score was 1. (E) The pStat3 expression score was 30.

    Article Snippet: For Glut1, the BenchMark XT was used.

    Techniques: Positron Emission Tomography, Expressing

    Western blot analysis of CD63, CD81, GPC-1, GLUT-1, and ADAM10 in the exosomes derived from the MDA and MCF cell lines. Full-length gels are shown in Figure S9 .

    Journal: Scientific Reports

    Article Title: The proteomic analysis of breast cell line exosomes reveals disease patterns and potential biomarkers

    doi: 10.1038/s41598-020-70393-4

    Figure Lengend Snippet: Western blot analysis of CD63, CD81, GPC-1, GLUT-1, and ADAM10 in the exosomes derived from the MDA and MCF cell lines. Full-length gels are shown in Figure S9 .

    Article Snippet: Flow cytometry (FC)For the validation of protein biomarkers, isolated exosomes were incubated for 40 min with one of the following primary antibodies: Glut-1 (Invitrogen, MA5-31960, SA0377), GLYP-1 (Invitrogen, PA5-86043, polyclonal) and ADAM10 (MyBioSource, MBS435195, polyclonal).

    Techniques: Western Blot, Gel Permeation Chromatography, Derivative Assay, Multiple Displacement Amplification

    tSNE plots of exosomes analyzed by FACS showing expression levels of ( a ) GPC-1 ( b ) GLUT-1 and ( c ) ADAM10 along with CD81 and CD63 on the surface of BC-derived exosomes.

    Journal: Scientific Reports

    Article Title: The proteomic analysis of breast cell line exosomes reveals disease patterns and potential biomarkers

    doi: 10.1038/s41598-020-70393-4

    Figure Lengend Snippet: tSNE plots of exosomes analyzed by FACS showing expression levels of ( a ) GPC-1 ( b ) GLUT-1 and ( c ) ADAM10 along with CD81 and CD63 on the surface of BC-derived exosomes.

    Article Snippet: Flow cytometry (FC)For the validation of protein biomarkers, isolated exosomes were incubated for 40 min with one of the following primary antibodies: Glut-1 (Invitrogen, MA5-31960, SA0377), GLYP-1 (Invitrogen, PA5-86043, polyclonal) and ADAM10 (MyBioSource, MBS435195, polyclonal).

    Techniques: FACS, Expressing, Gel Permeation Chromatography, Derivative Assay

    ( a–f ) Role of GLUT1 in VEGF-induced MC FN production. ( a ) Primary culture mouse MCs with the C57BL6 background were studied in culture with and without VEGF-A 165 peptide (2 ng/ml) for 48 h. VEGF-A 165 stimulated a two-fold increase in MC GLUT1

    Journal: Laboratory investigation; a journal of technical methods and pathology

    Article Title: Nephron-deficient Fvb mice develop rapidly progressive renal failure and heavy albuminuria involving excess glomerular GLUT1 and VEGF

    doi: 10.1038/labinvest.2009.95

    Figure Lengend Snippet: ( a–f ) Role of GLUT1 in VEGF-induced MC FN production. ( a ) Primary culture mouse MCs with the C57BL6 background were studied in culture with and without VEGF-A 165 peptide (2 ng/ml) for 48 h. VEGF-A 165 stimulated a two-fold increase in MC GLUT1

    Article Snippet: Both GLUT1 and VEGF were increased in MCs in vitro by mechanical stretch, and the increase in GLUT1 (48 h onward) preceded the increase in VEGF (72 h), suggesting that GLUT1 could act as a stimulus for the VEGF.

    Techniques:

    ( a–d ) Glomerular gene expression and glucose uptake rates in FvbROP Os/+ and FvbROP +/+ mice. ( a ) Western analyses of glomerular GLUT1, VEGF, PKCβ1, and PKCα in FvbROP +/+ and FvbROP Os/+ mice at 4 weeks of age ( n = 3–4

    Journal: Laboratory investigation; a journal of technical methods and pathology

    Article Title: Nephron-deficient Fvb mice develop rapidly progressive renal failure and heavy albuminuria involving excess glomerular GLUT1 and VEGF

    doi: 10.1038/labinvest.2009.95

    Figure Lengend Snippet: ( a–d ) Glomerular gene expression and glucose uptake rates in FvbROP Os/+ and FvbROP +/+ mice. ( a ) Western analyses of glomerular GLUT1, VEGF, PKCβ1, and PKCα in FvbROP +/+ and FvbROP Os/+ mice at 4 weeks of age ( n = 3–4

    Article Snippet: Both GLUT1 and VEGF were increased in MCs in vitro by mechanical stretch, and the increase in GLUT1 (48 h onward) preceded the increase in VEGF (72 h), suggesting that GLUT1 could act as a stimulus for the VEGF.

    Techniques: Expressing, Mouse Assay, Western Blot

    ( a–c ) Cyclic mechanical stretch induction of MC GLUT1 and VEGF expression. ( a ) GLUT1 protein levels were increased at 48 h of stretch, and remained elevated at 72 h (not shown). ( b ) VEGF protein expression was increased at 72h of stretch, but

    Journal: Laboratory investigation; a journal of technical methods and pathology

    Article Title: Nephron-deficient Fvb mice develop rapidly progressive renal failure and heavy albuminuria involving excess glomerular GLUT1 and VEGF

    doi: 10.1038/labinvest.2009.95

    Figure Lengend Snippet: ( a–c ) Cyclic mechanical stretch induction of MC GLUT1 and VEGF expression. ( a ) GLUT1 protein levels were increased at 48 h of stretch, and remained elevated at 72 h (not shown). ( b ) VEGF protein expression was increased at 72h of stretch, but

    Article Snippet: Both GLUT1 and VEGF were increased in MCs in vitro by mechanical stretch, and the increase in GLUT1 (48 h onward) preceded the increase in VEGF (72 h), suggesting that GLUT1 could act as a stimulus for the VEGF.

    Techniques: Expressing

    Effects of co-culture time and 3T3-Ll cell density on mRNA levels of Glutl and Glut4 in L6 cells. L6 cells were incubated with 3T3-Ll adipocytes at various times at 5 × 10 3 density and mRNA levels of (A) Glut1 and (B) Glut4 measured. L6 cells were incubated with 3T3-Ll adipocytes for 6 h at indicated densities and the mRNA levels of Glutl . P values by one-way ANOVA followed by post-hoc Tukey HSO test are shown. ANOVA P

    Journal: Frontiers in Endocrinology

    Article Title: Preliminary Evidence for Adipocytokine Signals in Skeletal Muscle Glucose Uptake

    doi: 10.3389/fendo.2018.00295

    Figure Lengend Snippet: Effects of co-culture time and 3T3-Ll cell density on mRNA levels of Glutl and Glut4 in L6 cells. L6 cells were incubated with 3T3-Ll adipocytes at various times at 5 × 10 3 density and mRNA levels of (A) Glut1 and (B) Glut4 measured. L6 cells were incubated with 3T3-Ll adipocytes for 6 h at indicated densities and the mRNA levels of Glutl . P values by one-way ANOVA followed by post-hoc Tukey HSO test are shown. ANOVA P

    Article Snippet: The Akt antibody and anti-phospho-specific Akt (Ser473 and Thr308) was purchased from Cell Signaling Technology (Boston, MA, USA) and anti-glucose transporter Glut1 antibody, from Abcam (Burlingame, CA, USA).

    Techniques: Co-Culture Assay, Incubation

    Effects of co-culture with 3T3-Ll cells and antioxidants on mRNA and protein levels of Glut1. (A) L6 cells were incubated with 3T3-Ll adipocytes or antioxidant Mn-TBAP (200 µM) for 24 h. mRNA levels of Glut1 were measured by quantitative real-time RT-PCR. Values are mean ± SD ( n = 5). P values by one-way ANOVA followed by post-hoc Tukey HSO test are shown. ANOVA P

    Journal: Frontiers in Endocrinology

    Article Title: Preliminary Evidence for Adipocytokine Signals in Skeletal Muscle Glucose Uptake

    doi: 10.3389/fendo.2018.00295

    Figure Lengend Snippet: Effects of co-culture with 3T3-Ll cells and antioxidants on mRNA and protein levels of Glut1. (A) L6 cells were incubated with 3T3-Ll adipocytes or antioxidant Mn-TBAP (200 µM) for 24 h. mRNA levels of Glut1 were measured by quantitative real-time RT-PCR. Values are mean ± SD ( n = 5). P values by one-way ANOVA followed by post-hoc Tukey HSO test are shown. ANOVA P

    Article Snippet: The Akt antibody and anti-phospho-specific Akt (Ser473 and Thr308) was purchased from Cell Signaling Technology (Boston, MA, USA) and anti-glucose transporter Glut1 antibody, from Abcam (Burlingame, CA, USA).

    Techniques: Co-Culture Assay, Incubation, Quantitative RT-PCR

    Representative image of immunostaining corresponding to the glucose transporter protein SLC2A1 (GLUT1) in 30 week-old Ins2 AKITA and WT cerebral cortex. Mural cells (ANPEP, green), glucose transporter 1 (GLUT1, red), endothelium (PECAM1, cyan). n = 2, scale bar 50 µm.

    Journal: Scientific Reports

    Article Title: Prolonged systemic hyperglycemia does not cause pericyte loss and permeability at the mouse blood-brain barrier

    doi: 10.1038/s41598-018-35576-0

    Figure Lengend Snippet: Representative image of immunostaining corresponding to the glucose transporter protein SLC2A1 (GLUT1) in 30 week-old Ins2 AKITA and WT cerebral cortex. Mural cells (ANPEP, green), glucose transporter 1 (GLUT1, red), endothelium (PECAM1, cyan). n = 2, scale bar 50 µm.

    Article Snippet: Primary antibodies used: goat anti-mouse PECAM1 (cat. #AF3628, R & D Systems, Abingdon, UK), rat anti-mouse Laminin α-2 chain (LAMA2) (cat. #Ab11576, Abcam, Cambridge, UK), rat anti-mouse ANPEP (cat. #MCA2183EL, AbD Serotec); rabbit anti-mouse Collagen IV (cat. #2150–1470, Bio-Rad Antibodies, Oxford, UK); rabbit anti-GLUT1 (cat. #07–1401, Millipore, Billerica, MA); rat anti-mouse PDGFR-β (cat. #14–1402, Thermo Fischer Scientific), rabbit anti-Vitronectin (VTN) (cat. #GWB-794F8F, Genway Biotech, San Diego, CA), rabbit anti-Desmin (DES) (cat. #Ab15200-1, Abcam), rat anti-GFAP (cat. #13-0300, Thermo Fischer Scientific), rabbit anti-AIF1 (cat. #019-19741, Wako, Neuss, Germany).

    Techniques: Immunostaining

    Intranasally delivered BMSC increased neurogenesis and angiogenesis in the ischemic Brain. A , B Animals were euthanized 14 days after stroke with and without BMSC treatment. Immunohistochemistry stained for BrdU (red), NeuN (green), Glut-1 (blue). Arrows point to the presence of co-labeled cells in the peri-infarct region 14 days post stroke. BrdU/NeuN co-labeled cells indicate the presence of proliferating neuronal cells. BrdU/Glut-1 co-labeled cells indicate the presence of proliferating blood vessel cells. Scale bars = 40 μm. C , D Enlarged image to show a colabeled NeuN and BrdU cell and the counting result of these cells is shown in the bar graph. There was a significant increase in the total number of NeuN/BrdU co-labeled cells in the BMSC treatment group compared to control. N = 5–6, p = 0.024; student’s t test. Scale bar = 10 μm. E , F Enlarged image shows Glut-1 and BrdU double positive endothelia cells. There was an increase in Glut-1/BrdU co-labeled cells in the HP-BMSC treatment compared to control. N = 5–6, *p

    Journal: BMC Neuroscience

    Article Title: Delayed and repeated intranasal delivery of bone marrow stromal cells increases regeneration and functional recovery after ischemic stroke in mice

    doi: 10.1186/s12868-018-0418-z

    Figure Lengend Snippet: Intranasally delivered BMSC increased neurogenesis and angiogenesis in the ischemic Brain. A , B Animals were euthanized 14 days after stroke with and without BMSC treatment. Immunohistochemistry stained for BrdU (red), NeuN (green), Glut-1 (blue). Arrows point to the presence of co-labeled cells in the peri-infarct region 14 days post stroke. BrdU/NeuN co-labeled cells indicate the presence of proliferating neuronal cells. BrdU/Glut-1 co-labeled cells indicate the presence of proliferating blood vessel cells. Scale bars = 40 μm. C , D Enlarged image to show a colabeled NeuN and BrdU cell and the counting result of these cells is shown in the bar graph. There was a significant increase in the total number of NeuN/BrdU co-labeled cells in the BMSC treatment group compared to control. N = 5–6, p = 0.024; student’s t test. Scale bar = 10 μm. E , F Enlarged image shows Glut-1 and BrdU double positive endothelia cells. There was an increase in Glut-1/BrdU co-labeled cells in the HP-BMSC treatment compared to control. N = 5–6, *p

    Article Snippet: Brain sections were blocked with 1% cold fish gelatin (Sigma) and incubated overnight at 4 °C with the following primary antibodies: Ms anti-NeuN (1:200; MAB377, Millipore, Billerica, MA), Rat anti-BrdU (1:400; AbD Serotec, Hercules, CA), and Rabbit anti-Glut-1 (Chemicon Millipore).

    Techniques: Immunohistochemistry, Staining, Labeling

    The image analysis results showed that the expression of GLUT1 on membrane was detected in cells transfected with cab39. Compared with un-transfected cells, the expression rates of GLUT1 in H460 cells (A) and LK2 cells (B) were 45.5% and 67.1%, respectively. Efficiency was measured by Western blotting. (C) The results of flow cytometry showed that the average density of glucose in H460 cells transfected with cab39 increased by 3.3 times ( p

    Journal: Therapeutic Advances in Chronic Disease

    Article Title: HPV16 E6/E7 promote the translocation and glucose uptake of GLUT1 by PI3K/AKT pathway via relieving miR-451 inhibitory effect on CAB39 in lung cancer cells

    doi: 10.1177/2040622320957143

    Figure Lengend Snippet: The image analysis results showed that the expression of GLUT1 on membrane was detected in cells transfected with cab39. Compared with un-transfected cells, the expression rates of GLUT1 in H460 cells (A) and LK2 cells (B) were 45.5% and 67.1%, respectively. Efficiency was measured by Western blotting. (C) The results of flow cytometry showed that the average density of glucose in H460 cells transfected with cab39 increased by 3.3 times ( p

    Article Snippet: Lee et al . demonstrated that protein kinase C involved in phosphorylation of GLUT1 can lead to a rapid increase in glucose uptake and enhanced cell surface localization of GLUT1 induced by 12-0-tetradecanoyl-phorbol-13-acetate (TPA).

    Techniques: Expressing, Transfection, Western Blot, Flow Cytometry

    HPV16 E6/E7 down-regulated the expression of miR-451 but up-regulated the expression of CAB39, PI3K (P85α), P-AKT (ser473), and GLUT1. Proteins of E6, E7, CAB39, PI3K (P85α), P-AKT (ser473), AKT, and GLUT1 were demonstrated by Western blotting and the mRNAs of E6, E7, miR-451, CAB39, and GLUT1 were demonstrated by qRT-PCR in H460 or A549 or LK2 cells. Mock, mock transfection or mock siRNA; NS, nonspecific siRNA; Vector, empty vector; qRT-PCR, quantitative real-time reverse transcriptase polymerase chain reaction. * p

    Journal: Therapeutic Advances in Chronic Disease

    Article Title: HPV16 E6/E7 promote the translocation and glucose uptake of GLUT1 by PI3K/AKT pathway via relieving miR-451 inhibitory effect on CAB39 in lung cancer cells

    doi: 10.1177/2040622320957143

    Figure Lengend Snippet: HPV16 E6/E7 down-regulated the expression of miR-451 but up-regulated the expression of CAB39, PI3K (P85α), P-AKT (ser473), and GLUT1. Proteins of E6, E7, CAB39, PI3K (P85α), P-AKT (ser473), AKT, and GLUT1 were demonstrated by Western blotting and the mRNAs of E6, E7, miR-451, CAB39, and GLUT1 were demonstrated by qRT-PCR in H460 or A549 or LK2 cells. Mock, mock transfection or mock siRNA; NS, nonspecific siRNA; Vector, empty vector; qRT-PCR, quantitative real-time reverse transcriptase polymerase chain reaction. * p

    Article Snippet: Lee et al . demonstrated that protein kinase C involved in phosphorylation of GLUT1 can lead to a rapid increase in glucose uptake and enhanced cell surface localization of GLUT1 induced by 12-0-tetradecanoyl-phorbol-13-acetate (TPA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Plasmid Preparation, Polymerase Chain Reaction

    CAB39 promoted the activation of GLUT1 and the promotion was dependent on the PI3K/AKT pathway. (A) A specific PI3K/AKT blocker, Miltefosine, inhibited the PI3K/AKT pathway in both H460 and LK2 cell lines transfected with cab39, the results showed that the effects of CAB39 on the expression of GLUT1 protein were reversed. (B) The results of image analysis showed that the promoting effects of CAB39 on GLUT1 translocation to the plasma membrane were reversed by confocal microscopy. (C) The promoting effects of CAB39 on the glucose uptake of GLUT1 were reversed by confocal microscopy. Vector, empty vector.

    Journal: Therapeutic Advances in Chronic Disease

    Article Title: HPV16 E6/E7 promote the translocation and glucose uptake of GLUT1 by PI3K/AKT pathway via relieving miR-451 inhibitory effect on CAB39 in lung cancer cells

    doi: 10.1177/2040622320957143

    Figure Lengend Snippet: CAB39 promoted the activation of GLUT1 and the promotion was dependent on the PI3K/AKT pathway. (A) A specific PI3K/AKT blocker, Miltefosine, inhibited the PI3K/AKT pathway in both H460 and LK2 cell lines transfected with cab39, the results showed that the effects of CAB39 on the expression of GLUT1 protein were reversed. (B) The results of image analysis showed that the promoting effects of CAB39 on GLUT1 translocation to the plasma membrane were reversed by confocal microscopy. (C) The promoting effects of CAB39 on the glucose uptake of GLUT1 were reversed by confocal microscopy. Vector, empty vector.

    Article Snippet: Lee et al . demonstrated that protein kinase C involved in phosphorylation of GLUT1 can lead to a rapid increase in glucose uptake and enhanced cell surface localization of GLUT1 induced by 12-0-tetradecanoyl-phorbol-13-acetate (TPA).

    Techniques: Activation Assay, Transfection, Expressing, Translocation Assay, Confocal Microscopy, Plasmid Preparation

    miR-451 down-regulated the expression of CAB39, PI3K (P85α), P-AKT (ser473), and GLUT1. Proteins of CAB39, PI3K (P85α), P-AKT (ser473), AKT, and GLUT1 were demonstrated by Western blotting and the mRNAs of miR-451, CAB39, and GLUT1 were demonstrated by qRT-PCR in H460 or A549 or LK2 cells. Mock, mock transfection or mock siRNA; NC, negative control; NS, nonspecific siRNA; Mimics, Hsa-miR-451 mimics; Inhibitor, miR-451 inhibitor; qRT-PCR, quantitative real-time reverse transcriptase polymerase chain reaction. * p

    Journal: Therapeutic Advances in Chronic Disease

    Article Title: HPV16 E6/E7 promote the translocation and glucose uptake of GLUT1 by PI3K/AKT pathway via relieving miR-451 inhibitory effect on CAB39 in lung cancer cells

    doi: 10.1177/2040622320957143

    Figure Lengend Snippet: miR-451 down-regulated the expression of CAB39, PI3K (P85α), P-AKT (ser473), and GLUT1. Proteins of CAB39, PI3K (P85α), P-AKT (ser473), AKT, and GLUT1 were demonstrated by Western blotting and the mRNAs of miR-451, CAB39, and GLUT1 were demonstrated by qRT-PCR in H460 or A549 or LK2 cells. Mock, mock transfection or mock siRNA; NC, negative control; NS, nonspecific siRNA; Mimics, Hsa-miR-451 mimics; Inhibitor, miR-451 inhibitor; qRT-PCR, quantitative real-time reverse transcriptase polymerase chain reaction. * p

    Article Snippet: Lee et al . demonstrated that protein kinase C involved in phosphorylation of GLUT1 can lead to a rapid increase in glucose uptake and enhanced cell surface localization of GLUT1 induced by 12-0-tetradecanoyl-phorbol-13-acetate (TPA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Negative Control, Polymerase Chain Reaction

    CAB39 up-regulated the expression of PI3K (P85α), P-AKT (ser473), and GLUT1. Proteins of CAB39, PI3K (P85α), P-AKT (ser473), AKT, and GLUT1 were demonstrated by Western blotting and the mRNAs of CAB39 and GLUT1 were demonstrated by qRT-PCR in H460 or A549 or LK2 cells. Mock, mock transfection or mock siRNA; NS, nonspecific siRNA; Vector, empty vector; ; qRT-PCR, quantitative real-time reverse transcriptase polymerase chain reaction. * p

    Journal: Therapeutic Advances in Chronic Disease

    Article Title: HPV16 E6/E7 promote the translocation and glucose uptake of GLUT1 by PI3K/AKT pathway via relieving miR-451 inhibitory effect on CAB39 in lung cancer cells

    doi: 10.1177/2040622320957143

    Figure Lengend Snippet: CAB39 up-regulated the expression of PI3K (P85α), P-AKT (ser473), and GLUT1. Proteins of CAB39, PI3K (P85α), P-AKT (ser473), AKT, and GLUT1 were demonstrated by Western blotting and the mRNAs of CAB39 and GLUT1 were demonstrated by qRT-PCR in H460 or A549 or LK2 cells. Mock, mock transfection or mock siRNA; NS, nonspecific siRNA; Vector, empty vector; ; qRT-PCR, quantitative real-time reverse transcriptase polymerase chain reaction. * p

    Article Snippet: Lee et al . demonstrated that protein kinase C involved in phosphorylation of GLUT1 can lead to a rapid increase in glucose uptake and enhanced cell surface localization of GLUT1 induced by 12-0-tetradecanoyl-phorbol-13-acetate (TPA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Plasmid Preparation, Polymerase Chain Reaction

    Insulin does not stimulate GLUT or IR protein expression in villous explants from term placentas of women with obesity. Villous explants from term placentas were treated with and without insulin, and (A) MVM GLUT1, (B) BM GLUT1 (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression was measured by western blot (n = 6 per group). Data are shown as mean ± SEM. Significance was evaluated with a paired t test.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Insulin Stimulates GLUT4 Trafficking to the Syncytiotrophoblast Basal Plasma Membrane in the Human Placenta

    doi: 10.1210/jc.2018-02778

    Figure Lengend Snippet: Insulin does not stimulate GLUT or IR protein expression in villous explants from term placentas of women with obesity. Villous explants from term placentas were treated with and without insulin, and (A) MVM GLUT1, (B) BM GLUT1 (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression was measured by western blot (n = 6 per group). Data are shown as mean ± SEM. Significance was evaluated with a paired t test.

    Article Snippet: Localization of GLUT4, GLUT1, and IR in term human placentaImmunohistochemistry was performed to determine the cellular localization of GLUT4, GLUT1, and IR in sections from term healthy placenta.

    Techniques: Expressing, Western Blot

    Insulin stimulates BM GLUT4 protein expression in villous explants from term placentas of women with normal BMI. Villous explants from term placentas were treated with and without insulin, and (A) MVM GLUT1, (B) BM GLUT1, (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression ( P = 0.0013) was measured by western blot (n = 7 to 8 per group). Data are shown as mean ± SEM. **P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Insulin Stimulates GLUT4 Trafficking to the Syncytiotrophoblast Basal Plasma Membrane in the Human Placenta

    doi: 10.1210/jc.2018-02778

    Figure Lengend Snippet: Insulin stimulates BM GLUT4 protein expression in villous explants from term placentas of women with normal BMI. Villous explants from term placentas were treated with and without insulin, and (A) MVM GLUT1, (B) BM GLUT1, (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression ( P = 0.0013) was measured by western blot (n = 7 to 8 per group). Data are shown as mean ± SEM. **P

    Article Snippet: Localization of GLUT4, GLUT1, and IR in term human placentaImmunohistochemistry was performed to determine the cellular localization of GLUT4, GLUT1, and IR in sections from term healthy placenta.

    Techniques: Expressing, Western Blot

    Effect of insulin on GLUT and IR protein expression in villous explants from early gestation placentas. Villous explants from early pregnancy placentas were treated with and without insulin and (A) MVM GLUT1, (B) BM GLUT1, (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression was determined by western blot (n = 6 to 10 per group). Data are shown as mean ± SEM. Significance was evaluated with a paired t test.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Insulin Stimulates GLUT4 Trafficking to the Syncytiotrophoblast Basal Plasma Membrane in the Human Placenta

    doi: 10.1210/jc.2018-02778

    Figure Lengend Snippet: Effect of insulin on GLUT and IR protein expression in villous explants from early gestation placentas. Villous explants from early pregnancy placentas were treated with and without insulin and (A) MVM GLUT1, (B) BM GLUT1, (C) MVM IR β , (D) BM IR β , and (E) BM GLUT4 protein expression was determined by western blot (n = 6 to 10 per group). Data are shown as mean ± SEM. Significance was evaluated with a paired t test.

    Article Snippet: Localization of GLUT4, GLUT1, and IR in term human placentaImmunohistochemistry was performed to determine the cellular localization of GLUT4, GLUT1, and IR in sections from term healthy placenta.

    Techniques: Expressing, Western Blot

    Glucose transporter protein expression in the MVM and BM in maternal obesity. GLUT4 protein expression in the BM and GLUT1 protein expression in the MVM and BM of placentas from women with normal BMI (Control, n = 11), women with obesity delivering AGA infants (n=12) or a macrosomic infant (Obese/Macro, n = 5), measured by Western blot, are shown. Data are shown as mean ± SEM. Each ANOVA contains three pairwise t tests and uses Bonferroni adjusted significance level α * = 0.05/3 = 0.0167. Groups with different superscript letters are significantly different at α * = 0.0167. (A) BM GLUT4 protein expression in Obese/Macro was significantly lower as compared with Obese ( P = 0.007) and Control ( P = 0.001) placentas; there was no significant difference in protein expression between Control and Obese placentas. (B) In the MVM GLUT1 protein expression was significantly lower in Obese/Macro as compared with Obese ( P = 0.0001) and Control placentas ( P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Insulin Stimulates GLUT4 Trafficking to the Syncytiotrophoblast Basal Plasma Membrane in the Human Placenta

    doi: 10.1210/jc.2018-02778

    Figure Lengend Snippet: Glucose transporter protein expression in the MVM and BM in maternal obesity. GLUT4 protein expression in the BM and GLUT1 protein expression in the MVM and BM of placentas from women with normal BMI (Control, n = 11), women with obesity delivering AGA infants (n=12) or a macrosomic infant (Obese/Macro, n = 5), measured by Western blot, are shown. Data are shown as mean ± SEM. Each ANOVA contains three pairwise t tests and uses Bonferroni adjusted significance level α * = 0.05/3 = 0.0167. Groups with different superscript letters are significantly different at α * = 0.0167. (A) BM GLUT4 protein expression in Obese/Macro was significantly lower as compared with Obese ( P = 0.007) and Control ( P = 0.001) placentas; there was no significant difference in protein expression between Control and Obese placentas. (B) In the MVM GLUT1 protein expression was significantly lower in Obese/Macro as compared with Obese ( P = 0.0001) and Control placentas ( P

    Article Snippet: Localization of GLUT4, GLUT1, and IR in term human placentaImmunohistochemistry was performed to determine the cellular localization of GLUT4, GLUT1, and IR in sections from term healthy placenta.

    Techniques: Expressing, Western Blot

    Cellular localization of GLUT4, GLUT1, and IR β . (A) GLUT4 (brown) is highly expressed in the BM of the syncytiotrophoblast and not in the MVM. (B) GLUT1 (brown) and (C) IR β (brown) are predominantly expressed in the MVM, with less expression in the BM. (D) Negative control without primary antibody added.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Insulin Stimulates GLUT4 Trafficking to the Syncytiotrophoblast Basal Plasma Membrane in the Human Placenta

    doi: 10.1210/jc.2018-02778

    Figure Lengend Snippet: Cellular localization of GLUT4, GLUT1, and IR β . (A) GLUT4 (brown) is highly expressed in the BM of the syncytiotrophoblast and not in the MVM. (B) GLUT1 (brown) and (C) IR β (brown) are predominantly expressed in the MVM, with less expression in the BM. (D) Negative control without primary antibody added.

    Article Snippet: Localization of GLUT4, GLUT1, and IR in term human placentaImmunohistochemistry was performed to determine the cellular localization of GLUT4, GLUT1, and IR in sections from term healthy placenta.

    Techniques: Expressing, Negative Control

    Glut1 expression increases GSK-3 phosphorylation, Mcl-1 level, and survival in primary hematopoietic cells. (A) Bone marrow cells were cultured in IL-3, infected with retrovirus to express Glut1 and truncated hNGFR as a marker on day 4, and stained by immunofluorescence for phospho-GSK-3 (red), hNGFR (green), and DAPI on day 7. (B to E) Glut1 protein levels were analyzed by immunoblotting (B), glucose uptake was measured (C), and phospho-GSK-3 levels were determined by immunoblotting (D) and quantified (E) in purified T lymphocytes from nontransgenic (Non) and Glut1-transgenic (Glut1) littermates freshly after purification or after 1 day of culture in the absence of stimulation (neglect). (F) Mcl-1 levels were determined by immunoblotting of freshly isolated, neglected, or GSK-3 inhibitor (10 μM SB216763 [SB])-treated nontransgenic T cells. Ratios are shown. (G) Mcl-1/actin ratio quantitated from nontransgenic and Glut1-transgenic T cells after isolation (fresh), neglect, or culture in GSK-3 inhibitor (SB). (H) Purified T cells from nontransgenic and Glut1-transgenic littermate mice were cultured without stimulation (neglect), and cell survival determined after 2 days. Values shown are the means from triplicate samples; error bars indicate standard deviations.

    Journal: Molecular and Cellular Biology

    Article Title: Glycogen Synthase Kinase 3? and 3? Mediate a Glucose-Sensitive Antiapoptotic Signaling Pathway To Stabilize Mcl-1 ▿

    doi: 10.1128/MCB.00153-07

    Figure Lengend Snippet: Glut1 expression increases GSK-3 phosphorylation, Mcl-1 level, and survival in primary hematopoietic cells. (A) Bone marrow cells were cultured in IL-3, infected with retrovirus to express Glut1 and truncated hNGFR as a marker on day 4, and stained by immunofluorescence for phospho-GSK-3 (red), hNGFR (green), and DAPI on day 7. (B to E) Glut1 protein levels were analyzed by immunoblotting (B), glucose uptake was measured (C), and phospho-GSK-3 levels were determined by immunoblotting (D) and quantified (E) in purified T lymphocytes from nontransgenic (Non) and Glut1-transgenic (Glut1) littermates freshly after purification or after 1 day of culture in the absence of stimulation (neglect). (F) Mcl-1 levels were determined by immunoblotting of freshly isolated, neglected, or GSK-3 inhibitor (10 μM SB216763 [SB])-treated nontransgenic T cells. Ratios are shown. (G) Mcl-1/actin ratio quantitated from nontransgenic and Glut1-transgenic T cells after isolation (fresh), neglect, or culture in GSK-3 inhibitor (SB). (H) Purified T cells from nontransgenic and Glut1-transgenic littermate mice were cultured without stimulation (neglect), and cell survival determined after 2 days. Values shown are the means from triplicate samples; error bars indicate standard deviations.

    Article Snippet: Antibodies used were rabbit anti-Bax (Cell Signaling), rabbit anti-Bim (BD Pharmingen), rabbit anti-Bcl-2 (BD Pharmingen), mouse anti-cytochrome c (BD Pharmingen), rabbit anti-Glut1 (Abcam), rabbit anti-phospho-GSK-3(Ser9/21) (Cell Signaling), rabbit anti-GSK-3β (Cell signaling), mouse anti-phospho-Akt (Ser473), rabbit anti-Mcl-1 (Santa Cruz and Biolegend), rabbit anti-Cox IV, rabbit antiubiquitin (Santa Cruz), rat anti-HA (Roche), rabbit anti-FLAG (Sigma), and mouse antiactin (Sigma).

    Techniques: Expressing, Cell Culture, Infection, Marker, Staining, Immunofluorescence, Purification, Transgenic Assay, Isolation, Mouse Assay

    GSK-3 phosphorylation is increased in Glut1- and HK1-expressing cells and regulates both Mcl-1 degradation and cell death. (A) Mcl-1-expressing control, Glut1, or Glut1/HK1 cells were analyzed for phospho-S159 and total Mcl-1. Ratios are shown. (B) Multiple FL5.12 clones expressing Glut1 and/or HK1 were analyzed for phospho-Akt, phospho-GSK-3, and total GSK-3β by immunoblotting. (C) Control and Glut1/HK1 FL5.12, 32D, and BAF3 cells were cultured in IL-3 or withdrawn from IL-3 for 6 hours, and phospho-GSK-3 and total GSK-3β were determined by immunoblotting. Ratios are shown. (D and E) Cells were treated with GSK-3 inhibitor (10 μM SB216763), followed by immunoblotting for Mcl-1 and actin (ratios are shown) (D) and determination of cell viability upon IL-3 withdrawal (E). (F and G) FL5.12 cells were transiently transfected with shRNAi for GFP or GSK-3β. Cells were analyzed by immunoblotting for GSK-3β, Mcl-1, and actin in the presence or absence of IL-3 (ratios are shown) (F), and cell survival upon IL-3 withdrawal was determined by PI exclusion (G). Values are means from triplicate samples; error bars indicate standard deviations. M, hMcl-1; N, Neo control; G, Glut1; H, HK1; GH, Glut1/HK1.

    Journal: Molecular and Cellular Biology

    Article Title: Glycogen Synthase Kinase 3? and 3? Mediate a Glucose-Sensitive Antiapoptotic Signaling Pathway To Stabilize Mcl-1 ▿

    doi: 10.1128/MCB.00153-07

    Figure Lengend Snippet: GSK-3 phosphorylation is increased in Glut1- and HK1-expressing cells and regulates both Mcl-1 degradation and cell death. (A) Mcl-1-expressing control, Glut1, or Glut1/HK1 cells were analyzed for phospho-S159 and total Mcl-1. Ratios are shown. (B) Multiple FL5.12 clones expressing Glut1 and/or HK1 were analyzed for phospho-Akt, phospho-GSK-3, and total GSK-3β by immunoblotting. (C) Control and Glut1/HK1 FL5.12, 32D, and BAF3 cells were cultured in IL-3 or withdrawn from IL-3 for 6 hours, and phospho-GSK-3 and total GSK-3β were determined by immunoblotting. Ratios are shown. (D and E) Cells were treated with GSK-3 inhibitor (10 μM SB216763), followed by immunoblotting for Mcl-1 and actin (ratios are shown) (D) and determination of cell viability upon IL-3 withdrawal (E). (F and G) FL5.12 cells were transiently transfected with shRNAi for GFP or GSK-3β. Cells were analyzed by immunoblotting for GSK-3β, Mcl-1, and actin in the presence or absence of IL-3 (ratios are shown) (F), and cell survival upon IL-3 withdrawal was determined by PI exclusion (G). Values are means from triplicate samples; error bars indicate standard deviations. M, hMcl-1; N, Neo control; G, Glut1; H, HK1; GH, Glut1/HK1.

    Article Snippet: Antibodies used were rabbit anti-Bax (Cell Signaling), rabbit anti-Bim (BD Pharmingen), rabbit anti-Bcl-2 (BD Pharmingen), mouse anti-cytochrome c (BD Pharmingen), rabbit anti-Glut1 (Abcam), rabbit anti-phospho-GSK-3(Ser9/21) (Cell Signaling), rabbit anti-GSK-3β (Cell signaling), mouse anti-phospho-Akt (Ser473), rabbit anti-Mcl-1 (Santa Cruz and Biolegend), rabbit anti-Cox IV, rabbit antiubiquitin (Santa Cruz), rat anti-HA (Roche), rabbit anti-FLAG (Sigma), and mouse antiactin (Sigma).

    Techniques: Expressing, Clone Assay, Cell Culture, Transfection

    Bim is necessary for death and is induced normally in Glut1/HK1 cells, yet Bim toxicity is inhibited. (A and B) Cells were transfected with control (GFP) or Bim shRNAi and analyzed in the presence or absence of IL-3 for Bim expression (A) and survival (B). (C and D) Bim RNA (C) and protein (D) induction after IL-3 withdrawal in control (Neo), Glut1/HK1, and Bcl-x L cells. Cells were cultured in the presence or absence of IL-3 for 10 h. Bim RNA levels were determined by RNase protection assay, and protein levels were determined by Western blotting. (E) Control (Neo), Glut1/HK1, and Bcl-x L cells were cultured in the presence or absence of IL-3 for the indicated times. Bim levels in mitochondrial fractions were determined by Western blotting. (F) Control (Neo) and Glut1/HK1 cells were cultured in the presence of or withdrawn from IL-3 for 10 h. Bim was immunoprecipitated, and the precipitates were analyzed by immunoblotting for Bim, Bcl-2, and Mcl-1. (G and H) Control (Neo) or Bim L plasmid was transiently transfected into control (Neo), Glut1/HK1, and Bcl-x L cells. Bim expression was determined by Western blotting (G), and cell viability was determined by PI exclusion (H). Values are means and standard deviations ( n = 3) (B and H). N, Neo control; GH, Glut1/HK1; X, Bcl-x L .

    Journal: Molecular and Cellular Biology

    Article Title: Glycogen Synthase Kinase 3? and 3? Mediate a Glucose-Sensitive Antiapoptotic Signaling Pathway To Stabilize Mcl-1 ▿

    doi: 10.1128/MCB.00153-07

    Figure Lengend Snippet: Bim is necessary for death and is induced normally in Glut1/HK1 cells, yet Bim toxicity is inhibited. (A and B) Cells were transfected with control (GFP) or Bim shRNAi and analyzed in the presence or absence of IL-3 for Bim expression (A) and survival (B). (C and D) Bim RNA (C) and protein (D) induction after IL-3 withdrawal in control (Neo), Glut1/HK1, and Bcl-x L cells. Cells were cultured in the presence or absence of IL-3 for 10 h. Bim RNA levels were determined by RNase protection assay, and protein levels were determined by Western blotting. (E) Control (Neo), Glut1/HK1, and Bcl-x L cells were cultured in the presence or absence of IL-3 for the indicated times. Bim levels in mitochondrial fractions were determined by Western blotting. (F) Control (Neo) and Glut1/HK1 cells were cultured in the presence of or withdrawn from IL-3 for 10 h. Bim was immunoprecipitated, and the precipitates were analyzed by immunoblotting for Bim, Bcl-2, and Mcl-1. (G and H) Control (Neo) or Bim L plasmid was transiently transfected into control (Neo), Glut1/HK1, and Bcl-x L cells. Bim expression was determined by Western blotting (G), and cell viability was determined by PI exclusion (H). Values are means and standard deviations ( n = 3) (B and H). N, Neo control; GH, Glut1/HK1; X, Bcl-x L .

    Article Snippet: Antibodies used were rabbit anti-Bax (Cell Signaling), rabbit anti-Bim (BD Pharmingen), rabbit anti-Bcl-2 (BD Pharmingen), mouse anti-cytochrome c (BD Pharmingen), rabbit anti-Glut1 (Abcam), rabbit anti-phospho-GSK-3(Ser9/21) (Cell Signaling), rabbit anti-GSK-3β (Cell signaling), mouse anti-phospho-Akt (Ser473), rabbit anti-Mcl-1 (Santa Cruz and Biolegend), rabbit anti-Cox IV, rabbit antiubiquitin (Santa Cruz), rat anti-HA (Roche), rabbit anti-FLAG (Sigma), and mouse antiactin (Sigma).

    Techniques: Transfection, Expressing, Cell Culture, Rnase Protection Assay, Western Blot, Immunoprecipitation, Plasmid Preparation

    Glut1/HK1 expression increases glucose metabolism and delays commitment to cell death. (A) Glycolytic flux was measured in control (Neo) and Glut1/HK1 cells in the presence or absence of IL-3. (B) Tandem mass spectrometry-based analysis of organic acids in control and Glut1/HK1 cells cultured in the presence or absence of IL-3 for 6 h. (C) Clonogenicity of multiple individual control, Glut1/HK1, and Bcl-x L cells cultured in the absence of IL-3 for the indicated times, followed by readdition of IL-3. (D) Control and Glut1/HK1 cells were IL-3 withdrawn and cultured in 10 mM glucose, 10 mM 2DOG plus 1 mM methylpyruvate, or 5 mM glucose plus 5 mM 2DOG, and viability was determined by PI exclusion. (E) Bax activation was analyzed by immunoprecipitation (I.P.) of active Bax from cells cultured in the presence or absence of IL-3 for 10 h followed by Western blotting for total Bax. (F) Cytochrome c release in control (Neo), Glut1/HK1, and Bcl-x L cells after IL-3 withdrawal. Cells were cultured in the presence or absence of IL-3 for the indicated times, fractionated to isolate cytosolic and mitochondrial proteins, and analyzed by Western blotting for cytochrome c . Values are means and standard deviation ( n = 3) (A, B, and D) or means and standard errors of the means ( n = 5) (C). N, Neo control; GH, Glut1/HK1; X, Bcl-x L .

    Journal: Molecular and Cellular Biology

    Article Title: Glycogen Synthase Kinase 3? and 3? Mediate a Glucose-Sensitive Antiapoptotic Signaling Pathway To Stabilize Mcl-1 ▿

    doi: 10.1128/MCB.00153-07

    Figure Lengend Snippet: Glut1/HK1 expression increases glucose metabolism and delays commitment to cell death. (A) Glycolytic flux was measured in control (Neo) and Glut1/HK1 cells in the presence or absence of IL-3. (B) Tandem mass spectrometry-based analysis of organic acids in control and Glut1/HK1 cells cultured in the presence or absence of IL-3 for 6 h. (C) Clonogenicity of multiple individual control, Glut1/HK1, and Bcl-x L cells cultured in the absence of IL-3 for the indicated times, followed by readdition of IL-3. (D) Control and Glut1/HK1 cells were IL-3 withdrawn and cultured in 10 mM glucose, 10 mM 2DOG plus 1 mM methylpyruvate, or 5 mM glucose plus 5 mM 2DOG, and viability was determined by PI exclusion. (E) Bax activation was analyzed by immunoprecipitation (I.P.) of active Bax from cells cultured in the presence or absence of IL-3 for 10 h followed by Western blotting for total Bax. (F) Cytochrome c release in control (Neo), Glut1/HK1, and Bcl-x L cells after IL-3 withdrawal. Cells were cultured in the presence or absence of IL-3 for the indicated times, fractionated to isolate cytosolic and mitochondrial proteins, and analyzed by Western blotting for cytochrome c . Values are means and standard deviation ( n = 3) (A, B, and D) or means and standard errors of the means ( n = 5) (C). N, Neo control; GH, Glut1/HK1; X, Bcl-x L .

    Article Snippet: Antibodies used were rabbit anti-Bax (Cell Signaling), rabbit anti-Bim (BD Pharmingen), rabbit anti-Bcl-2 (BD Pharmingen), mouse anti-cytochrome c (BD Pharmingen), rabbit anti-Glut1 (Abcam), rabbit anti-phospho-GSK-3(Ser9/21) (Cell Signaling), rabbit anti-GSK-3β (Cell signaling), mouse anti-phospho-Akt (Ser473), rabbit anti-Mcl-1 (Santa Cruz and Biolegend), rabbit anti-Cox IV, rabbit antiubiquitin (Santa Cruz), rat anti-HA (Roche), rabbit anti-FLAG (Sigma), and mouse antiactin (Sigma).

    Techniques: Expressing, Mass Spectrometry, Cell Culture, Activation Assay, Immunoprecipitation, Western Blot, Standard Deviation

    Maintenance of Mcl-1 protein level is critical for protection of Glut1/HK1 cells upon IL-3 withdrawal. (A and B) Control, Glut1/HK1, and Bcl-x L cells were cultured in the presence or absence of IL-3 for 10 h, and mitochondria were isolated. (A and B) Representative gel (A) and quantitated averages and standard deviations of mitochondrial Mcl-1 levels using Bcl-2 as a normalization control (B). (C) Control (Neo), Glut1/HK1, and Bcl-x L cells were withdrawn from IL-3, and the total cellular Mcl-1/actin ratio was determined over time. (D and E) Control and Glut1/HK1 cells were transiently transfected with control (GFP) or Mcl-1 shRNAi, and then Mcl-1 protein level was determined by immunoblotting (D) and cell viability was observed by PI exclusion (E). Values are means for cell survival from triplicate samples; error bars indicate standard deviations. N, Neo control; GH, Glut1/HK1; X, Bcl-x L .

    Journal: Molecular and Cellular Biology

    Article Title: Glycogen Synthase Kinase 3? and 3? Mediate a Glucose-Sensitive Antiapoptotic Signaling Pathway To Stabilize Mcl-1 ▿

    doi: 10.1128/MCB.00153-07

    Figure Lengend Snippet: Maintenance of Mcl-1 protein level is critical for protection of Glut1/HK1 cells upon IL-3 withdrawal. (A and B) Control, Glut1/HK1, and Bcl-x L cells were cultured in the presence or absence of IL-3 for 10 h, and mitochondria were isolated. (A and B) Representative gel (A) and quantitated averages and standard deviations of mitochondrial Mcl-1 levels using Bcl-2 as a normalization control (B). (C) Control (Neo), Glut1/HK1, and Bcl-x L cells were withdrawn from IL-3, and the total cellular Mcl-1/actin ratio was determined over time. (D and E) Control and Glut1/HK1 cells were transiently transfected with control (GFP) or Mcl-1 shRNAi, and then Mcl-1 protein level was determined by immunoblotting (D) and cell viability was observed by PI exclusion (E). Values are means for cell survival from triplicate samples; error bars indicate standard deviations. N, Neo control; GH, Glut1/HK1; X, Bcl-x L .

    Article Snippet: Antibodies used were rabbit anti-Bax (Cell Signaling), rabbit anti-Bim (BD Pharmingen), rabbit anti-Bcl-2 (BD Pharmingen), mouse anti-cytochrome c (BD Pharmingen), rabbit anti-Glut1 (Abcam), rabbit anti-phospho-GSK-3(Ser9/21) (Cell Signaling), rabbit anti-GSK-3β (Cell signaling), mouse anti-phospho-Akt (Ser473), rabbit anti-Mcl-1 (Santa Cruz and Biolegend), rabbit anti-Cox IV, rabbit antiubiquitin (Santa Cruz), rat anti-HA (Roche), rabbit anti-FLAG (Sigma), and mouse antiactin (Sigma).

    Techniques: Cell Culture, Isolation, Transfection

    Glucose uptake attenuates Mcl-1 ubiquitination and degradation after growth factor withdrawal. (A, B, and C) Control and Glut1/HK1 cells were withdrawn from IL-3 for 8 h and then treated with cycloheximide (CHX) to observe Mcl-1 degradation. (A) Total cellular Mcl-1 as a fraction of starting levels prior to CHX treatment. (B and C) Representative gels (B) and quantitated averages and standard deviations from three independent experiments (C) of mitochondrial Mcl-1 levels, using Bcl-2 as a normalization control, before (−) and after (+) CHX treatment. (D) HA-Mcl-1-expressing control and Glut1/HK1 cells were transiently transfected with FLAG-ubiquitin (FLAG-Ub) and withdrawn from IL-3 for 10 h. The proteasome inhibitor MG-132 (20 μM) was added in the last hour of culture. Lysates were immunoprecipitated with anti-HA and probed with anti-FLAG, anti-HA, or anti-heavy chain immunoglobulin. N, Neo control; GH, Glut1/HK1.

    Journal: Molecular and Cellular Biology

    Article Title: Glycogen Synthase Kinase 3? and 3? Mediate a Glucose-Sensitive Antiapoptotic Signaling Pathway To Stabilize Mcl-1 ▿

    doi: 10.1128/MCB.00153-07

    Figure Lengend Snippet: Glucose uptake attenuates Mcl-1 ubiquitination and degradation after growth factor withdrawal. (A, B, and C) Control and Glut1/HK1 cells were withdrawn from IL-3 for 8 h and then treated with cycloheximide (CHX) to observe Mcl-1 degradation. (A) Total cellular Mcl-1 as a fraction of starting levels prior to CHX treatment. (B and C) Representative gels (B) and quantitated averages and standard deviations from three independent experiments (C) of mitochondrial Mcl-1 levels, using Bcl-2 as a normalization control, before (−) and after (+) CHX treatment. (D) HA-Mcl-1-expressing control and Glut1/HK1 cells were transiently transfected with FLAG-ubiquitin (FLAG-Ub) and withdrawn from IL-3 for 10 h. The proteasome inhibitor MG-132 (20 μM) was added in the last hour of culture. Lysates were immunoprecipitated with anti-HA and probed with anti-FLAG, anti-HA, or anti-heavy chain immunoglobulin. N, Neo control; GH, Glut1/HK1.

    Article Snippet: Antibodies used were rabbit anti-Bax (Cell Signaling), rabbit anti-Bim (BD Pharmingen), rabbit anti-Bcl-2 (BD Pharmingen), mouse anti-cytochrome c (BD Pharmingen), rabbit anti-Glut1 (Abcam), rabbit anti-phospho-GSK-3(Ser9/21) (Cell Signaling), rabbit anti-GSK-3β (Cell signaling), mouse anti-phospho-Akt (Ser473), rabbit anti-Mcl-1 (Santa Cruz and Biolegend), rabbit anti-Cox IV, rabbit antiubiquitin (Santa Cruz), rat anti-HA (Roche), rabbit anti-FLAG (Sigma), and mouse antiactin (Sigma).

    Techniques: Expressing, Transfection, Immunoprecipitation

    HOXA9 represses the expression of HIF-1α and its downstream glycolytic genes by directly binding to the promoter regions. a , b The mRNA or protein expression of HOXA9, HIF-1α, HK2, GLUT1, and PDK1 was detected by qRT-PCR or western blot, respectively, in A431 cells after depletion of HOXA9 by two siRNAs or overexpression of HOXA9. The qRT-PCR data were normalized to GAPDH gene expression. In western blots, GAPDH was used as a loading control. c Known or predicted binding sites of HOXA9 (diamond) or HRE elements for HIF-1α (delta) in the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 ). d Electrophoretic mobility shift assay (EMSA) was used to detect the direct association of purified HOXA9 protein with its binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 . The black arrows indicate the binding complex of the HOXA9 protein with the probe containing the known or predicted binding sites whereas the white arrows indicate the supershift generated by the association of the anti-HOXA9 antibody with the above complexes. Also, the specificity of the direct associations was supported by the loss of binding activities using mutated probes. e The binding enrichment of HOXA9 at the above binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 was detected by ChIP-PCR after knockdown of HOXA9. f The binding enrichment of HIF-1α at the above binding sites on the promoter regions was detected after overexpression of HOXA9. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P

    Journal: Nature Communications

    Article Title: HOXA9 inhibits HIF-1α-mediated glycolysis through interacting with CRIP2 to repress cutaneous squamous cell carcinoma development

    doi: 10.1038/s41467-018-03914-5

    Figure Lengend Snippet: HOXA9 represses the expression of HIF-1α and its downstream glycolytic genes by directly binding to the promoter regions. a , b The mRNA or protein expression of HOXA9, HIF-1α, HK2, GLUT1, and PDK1 was detected by qRT-PCR or western blot, respectively, in A431 cells after depletion of HOXA9 by two siRNAs or overexpression of HOXA9. The qRT-PCR data were normalized to GAPDH gene expression. In western blots, GAPDH was used as a loading control. c Known or predicted binding sites of HOXA9 (diamond) or HRE elements for HIF-1α (delta) in the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 ). d Electrophoretic mobility shift assay (EMSA) was used to detect the direct association of purified HOXA9 protein with its binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 . The black arrows indicate the binding complex of the HOXA9 protein with the probe containing the known or predicted binding sites whereas the white arrows indicate the supershift generated by the association of the anti-HOXA9 antibody with the above complexes. Also, the specificity of the direct associations was supported by the loss of binding activities using mutated probes. e The binding enrichment of HOXA9 at the above binding sites on the promoter regions of HIF1A , HK2 , GLUT1 , and PDK1 was detected by ChIP-PCR after knockdown of HOXA9. f The binding enrichment of HIF-1α at the above binding sites on the promoter regions was detected after overexpression of HOXA9. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P

    Article Snippet: The following primary antibodies and dilutions were used: HOXA9 (Abcam, ab140631, 1:2000) and CRIP2 (Abcam, ab151496, 1:2000); HIF-1α (Santa Cruz Biotechnology, sc-10790, 1:2000), HK2 (Santa Cruz Biotechnology, sc-374091, 1:2000), GLUT1 (Cell Signaling Technology, 12939S, 1:2000), and PDK1 (Cell Signaling Technology, 3062, 1:2000); and α-TUBULIN (Santa Cruz Biotechnology, sc-5286, 1:2000) and GAPDH (Santa Cruz Biotechnology, sc-25778, 1:5000).

    Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Western Blot, Over Expression, Electrophoretic Mobility Shift Assay, Purification, Generated, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    Loss of HOXA9 leads to enhanced glycolysis and tumor growth in vivo. siNC and siHOXA9 oligos were injected into A431 cell xenografts every 3 days. a Loss of HOXA9 promotes subcutaneous tumor growth in a mouse xenograft model. Tumor volumes (mm 3 ) were plotted according to day. Tumor volume statistical data represent the average of four independent experiments ± s.d, respectively. b The mice were sacrificed at the end of the experiment and images taken along with the dissected tumors from five representative mice are shown. White arrows indicate the siNC-treated xenografts whereas black arrows indicate siHOXA9-treated xenografts. Scale bar, 1 cm. c The expression of HOXA9 , HIF1A , HK2 , GLUT1 , and PDK1 was measured in the dissected tumors by qRT-PCR. qRT-PCR statistical data represent the average of four independent experiments ± s.d. d The protein expression of HOXA9, HIF-1α, HK2, GLUT1, and PDK1 was detected in xenografts after siHOXA9 treatment by western blot. e Histopathology analysis (IHC staining) of HOXA9, HIF-1α, HK2, GLUT1, and PDK1 on tumor sections. HOXA9 pre-absorption tests was also performed to validate the specificity of HOXA9 antibody. Scale bar, 100 µm (200×). f Comparison of glucose consumption between siHOXA9-treated and siNC-treated xenograft tumors by microPET/CT imaging of the uptake and retention of 18 F-FDG injected via the tail vein. A representative microPET/CT image is shown. g A model of the miR-365-HOXA9-HIF-1α glycolysis-regulatory axis in cSCC development. In cSCC tumors, loss of HOXA9 up-regulates HIF-1α and its downstream glycolytic genes of HK2 , GLUT1 , and PDK1 in the HIF-1 pathway, which contributes to the enhancement of glycolysis and promotes cSCC progression. In normal skin or HOXA9-treated cSCC, HOXA9 interacts with CRIP2 and epigenetically represses HIF-1α expression, which leads to the replacement of HIF-1α by the HOXA9-CRIP2 complex at the promoter regions and represses the expression of glycolytic genes including HK2 , GLUT1 , and PDK1 , which subsequently contributes to the inhibition of tumor progression. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P

    Journal: Nature Communications

    Article Title: HOXA9 inhibits HIF-1α-mediated glycolysis through interacting with CRIP2 to repress cutaneous squamous cell carcinoma development

    doi: 10.1038/s41467-018-03914-5

    Figure Lengend Snippet: Loss of HOXA9 leads to enhanced glycolysis and tumor growth in vivo. siNC and siHOXA9 oligos were injected into A431 cell xenografts every 3 days. a Loss of HOXA9 promotes subcutaneous tumor growth in a mouse xenograft model. Tumor volumes (mm 3 ) were plotted according to day. Tumor volume statistical data represent the average of four independent experiments ± s.d, respectively. b The mice were sacrificed at the end of the experiment and images taken along with the dissected tumors from five representative mice are shown. White arrows indicate the siNC-treated xenografts whereas black arrows indicate siHOXA9-treated xenografts. Scale bar, 1 cm. c The expression of HOXA9 , HIF1A , HK2 , GLUT1 , and PDK1 was measured in the dissected tumors by qRT-PCR. qRT-PCR statistical data represent the average of four independent experiments ± s.d. d The protein expression of HOXA9, HIF-1α, HK2, GLUT1, and PDK1 was detected in xenografts after siHOXA9 treatment by western blot. e Histopathology analysis (IHC staining) of HOXA9, HIF-1α, HK2, GLUT1, and PDK1 on tumor sections. HOXA9 pre-absorption tests was also performed to validate the specificity of HOXA9 antibody. Scale bar, 100 µm (200×). f Comparison of glucose consumption between siHOXA9-treated and siNC-treated xenograft tumors by microPET/CT imaging of the uptake and retention of 18 F-FDG injected via the tail vein. A representative microPET/CT image is shown. g A model of the miR-365-HOXA9-HIF-1α glycolysis-regulatory axis in cSCC development. In cSCC tumors, loss of HOXA9 up-regulates HIF-1α and its downstream glycolytic genes of HK2 , GLUT1 , and PDK1 in the HIF-1 pathway, which contributes to the enhancement of glycolysis and promotes cSCC progression. In normal skin or HOXA9-treated cSCC, HOXA9 interacts with CRIP2 and epigenetically represses HIF-1α expression, which leads to the replacement of HIF-1α by the HOXA9-CRIP2 complex at the promoter regions and represses the expression of glycolytic genes including HK2 , GLUT1 , and PDK1 , which subsequently contributes to the inhibition of tumor progression. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P

    Article Snippet: The following primary antibodies and dilutions were used: HOXA9 (Abcam, ab140631, 1:2000) and CRIP2 (Abcam, ab151496, 1:2000); HIF-1α (Santa Cruz Biotechnology, sc-10790, 1:2000), HK2 (Santa Cruz Biotechnology, sc-374091, 1:2000), GLUT1 (Cell Signaling Technology, 12939S, 1:2000), and PDK1 (Cell Signaling Technology, 3062, 1:2000); and α-TUBULIN (Santa Cruz Biotechnology, sc-5286, 1:2000) and GAPDH (Santa Cruz Biotechnology, sc-25778, 1:5000).

    Techniques: In Vivo, Injection, Mouse Assay, Expressing, Quantitative RT-PCR, Western Blot, Histopathology, Immunohistochemistry, Staining, Imaging, Inhibition

    The glycolysis-inhibitory role of HOXA9 is dependent on CRIP2. a Western blot detection showed that CRIP2 knockdown led to significant upregulation of HIF-1α, HK2, GLUT1, and PDK1, whereas HOXA9 expression was not affected. b – d Depletion of CRIP2 enhanced cell proliferation, migration, and invasiveness of A431 cells by CCK-8 assay, transwell migration assay, and Matrigel invasiveness measurement. e , f ECAR and OCR analysis of A431 cells following depletion of CRIP2 by siRNA as summarized from raw data. g Western blot detection showing the variations of protein expression levels of HOXA9, CRIP2, HIF-1α, HK2, GLUT1, and PDK1 in response to CRIP2 knockdown and/or HOXA9 overexpression. h , i ECAR and OCR analysis of A431 cells depleted of CRIP2 by siRNA and/or overexpressing HOXA9 as summarized from raw data. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P

    Journal: Nature Communications

    Article Title: HOXA9 inhibits HIF-1α-mediated glycolysis through interacting with CRIP2 to repress cutaneous squamous cell carcinoma development

    doi: 10.1038/s41467-018-03914-5

    Figure Lengend Snippet: The glycolysis-inhibitory role of HOXA9 is dependent on CRIP2. a Western blot detection showed that CRIP2 knockdown led to significant upregulation of HIF-1α, HK2, GLUT1, and PDK1, whereas HOXA9 expression was not affected. b – d Depletion of CRIP2 enhanced cell proliferation, migration, and invasiveness of A431 cells by CCK-8 assay, transwell migration assay, and Matrigel invasiveness measurement. e , f ECAR and OCR analysis of A431 cells following depletion of CRIP2 by siRNA as summarized from raw data. g Western blot detection showing the variations of protein expression levels of HOXA9, CRIP2, HIF-1α, HK2, GLUT1, and PDK1 in response to CRIP2 knockdown and/or HOXA9 overexpression. h , i ECAR and OCR analysis of A431 cells depleted of CRIP2 by siRNA and/or overexpressing HOXA9 as summarized from raw data. Each experiment was performed in triplicate and data are presented as mean ± s.d. One-Way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (* P

    Article Snippet: The following primary antibodies and dilutions were used: HOXA9 (Abcam, ab140631, 1:2000) and CRIP2 (Abcam, ab151496, 1:2000); HIF-1α (Santa Cruz Biotechnology, sc-10790, 1:2000), HK2 (Santa Cruz Biotechnology, sc-374091, 1:2000), GLUT1 (Cell Signaling Technology, 12939S, 1:2000), and PDK1 (Cell Signaling Technology, 3062, 1:2000); and α-TUBULIN (Santa Cruz Biotechnology, sc-5286, 1:2000) and GAPDH (Santa Cruz Biotechnology, sc-25778, 1:5000).

    Techniques: Western Blot, Expressing, Migration, CCK-8 Assay, Transwell Migration Assay, Over Expression

    Graphical representation of PrP C -mediated modulation of glucose homeostasis through iron. (1 2) PrP C facilitates cellular uptake of Tf-Fe 3+ and non-Tf-bound iron (Fe 3+ . (3) , resulting in the downregulation of GLUT2 in pancreatic β-cells and hepatocytes (black), GLUT 3 in neuronal cells (red), and GLUT1 at the blood-retinal barrier (blue). (4) Reduced uptake of glucose downregulates insulin, resulting in hyperglycemia. (5) Down-regulation or deletion of PrP C and glucose transporters. (6) Increased uptake of glucose stimulates insulin secretion with resultant hypoglycemia.

    Journal: Scientific Reports

    Article Title: Prion protein modulates glucose homeostasis by altering intracellular iron

    doi: 10.1038/s41598-018-24786-1

    Figure Lengend Snippet: Graphical representation of PrP C -mediated modulation of glucose homeostasis through iron. (1 2) PrP C facilitates cellular uptake of Tf-Fe 3+ and non-Tf-bound iron (Fe 3+ . (3) , resulting in the downregulation of GLUT2 in pancreatic β-cells and hepatocytes (black), GLUT 3 in neuronal cells (red), and GLUT1 at the blood-retinal barrier (blue). (4) Reduced uptake of glucose downregulates insulin, resulting in hyperglycemia. (5) Down-regulation or deletion of PrP C and glucose transporters. (6) Increased uptake of glucose stimulates insulin secretion with resultant hypoglycemia.

    Article Snippet: Other antibodies were obtained from the following sources: ferritin specific for heavy and light chain (F5012) from Sigma Aldrich, USA, GLUT-1 (NB110-39113) from Novus Biologicals, GLUT-2 (ab54460), and GLUT-3 (ab41525) from Abcam, USA, TfR (13-6800) from Invitrogen, USA, insulin from Santa Cruz Biotechnology Inc. (sc-9168) and Novus Biologicals (NBP2-34260), USA (recognize insulin and a 51-amino acid polypeptide composed of A and B chains connected through the C-peptide), glucagon (sc-13091) from Santa Cruz Biotechnology Inc, USA, and β-actin (MAB1501) from Millipore, USA.

    Techniques:

    CD4+Glut1+ T cell percentage inversely correlates with CD4+ T cell counts (A,B) , CD4+ T cell percentage (C,D) , and CD4/CD8 ratio (E,F) ). cART, combination antiretroviral therapy.

    Journal: Frontiers in Immunology

    Article Title: Polymorphism rs1385129 Within Glut1 Gene SLC2A1 Is Linked to Poor CD4+ T Cell Recovery in Antiretroviral-Treated HIV+ Individuals

    doi: 10.3389/fimmu.2018.00900

    Figure Lengend Snippet: CD4+Glut1+ T cell percentage inversely correlates with CD4+ T cell counts (A,B) , CD4+ T cell percentage (C,D) , and CD4/CD8 ratio (E,F) ). cART, combination antiretroviral therapy.

    Article Snippet: The Glut1 antibodies [FAB1418A and MAB1418 clones (R & D Systems, Minneapolis, MN, USA)] conjugated with APC and FITC were used to analyze Glut1 on CD4+ and CD8+ T cells using the staining procedure previously described ( ).

    Techniques:

    Gaiting strategy and analysis of CD4+Glut1+ T cell percentage in HIV-negative and HIV-positive populations. (A–C) Percentage of Glut1 expressing CD3+CD4+ T cells within the lymphocyte population were defined using side scatter (SSC) and the Glut1 fluorophore (FITC); (D,E) comparison between HIV-negative controls and HIV-positive treatment-naïve groups with favorable and non-favorable disease progression; (F,G) comparison between HIV-negative controls and HIV+/cART responder and non-responder groups. Horizontal black lines refer to median CD4+Glut1+ or CD8+Glut1+ T cell percentages of each group. cART, combination antiretroviral therapy.

    Journal: Frontiers in Immunology

    Article Title: Polymorphism rs1385129 Within Glut1 Gene SLC2A1 Is Linked to Poor CD4+ T Cell Recovery in Antiretroviral-Treated HIV+ Individuals

    doi: 10.3389/fimmu.2018.00900

    Figure Lengend Snippet: Gaiting strategy and analysis of CD4+Glut1+ T cell percentage in HIV-negative and HIV-positive populations. (A–C) Percentage of Glut1 expressing CD3+CD4+ T cells within the lymphocyte population were defined using side scatter (SSC) and the Glut1 fluorophore (FITC); (D,E) comparison between HIV-negative controls and HIV-positive treatment-naïve groups with favorable and non-favorable disease progression; (F,G) comparison between HIV-negative controls and HIV+/cART responder and non-responder groups. Horizontal black lines refer to median CD4+Glut1+ or CD8+Glut1+ T cell percentages of each group. cART, combination antiretroviral therapy.

    Article Snippet: The Glut1 antibodies [FAB1418A and MAB1418 clones (R & D Systems, Minneapolis, MN, USA)] conjugated with APC and FITC were used to analyze Glut1 on CD4+ and CD8+ T cells using the staining procedure previously described ( ).

    Techniques: Expressing

    Synovial tissue (ST) angiogenesis, oxidative stress and cellular bioenergetics. To support the concept that oxidative stress, angiogenesis and energy metabolism are interconnected processes that co-exist during the inflammation milieu, double-immunofluorescence staining was performed. ST slides were co-incubated with primary mouse antibody against human 4-hydroxy-2-nonenal (4-HNE) and with primary rabbit antibodies against angiogenic factors (vascular endothelial growth factor [VEGF], angiopoietin 2 [Ang2], tyrosine kinase receptor [Tie2]), glycolytic proteins (glyceraldehyde 3-phosphate dehydrogenase [GAPDH], pyruvate kinase isozyme 2 [PKM2], glucose transporter 1 [GLUT1]) and a mitochondrial marker (adenosine triphosphate synthase subunit β [ATP5B]). Representative merged immunofluorescence images demonstrate examples of co-localisation ( yellow ) of 4-HNE with VEGF, Ang2, Tie2, GAPDH, PKM2, GLUT1 and ATP5B. Cells stained green are positive for 4-HNE only; cells stained red are positive only for VEGF, Ang2, Tie2, GAPDH, PKM2, GLUT1 and ATP5B. Arrows indicate examples of co-localisation. Magnification of photomicrographs × 20, insets : Figure S4

    Journal: Arthritis Research & Therapy

    Article Title: Oxidative stress impairs energy metabolism in primary cells and synovial tissue of patients with rheumatoid arthritis

    doi: 10.1186/s13075-018-1592-1

    Figure Lengend Snippet: Synovial tissue (ST) angiogenesis, oxidative stress and cellular bioenergetics. To support the concept that oxidative stress, angiogenesis and energy metabolism are interconnected processes that co-exist during the inflammation milieu, double-immunofluorescence staining was performed. ST slides were co-incubated with primary mouse antibody against human 4-hydroxy-2-nonenal (4-HNE) and with primary rabbit antibodies against angiogenic factors (vascular endothelial growth factor [VEGF], angiopoietin 2 [Ang2], tyrosine kinase receptor [Tie2]), glycolytic proteins (glyceraldehyde 3-phosphate dehydrogenase [GAPDH], pyruvate kinase isozyme 2 [PKM2], glucose transporter 1 [GLUT1]) and a mitochondrial marker (adenosine triphosphate synthase subunit β [ATP5B]). Representative merged immunofluorescence images demonstrate examples of co-localisation ( yellow ) of 4-HNE with VEGF, Ang2, Tie2, GAPDH, PKM2, GLUT1 and ATP5B. Cells stained green are positive for 4-HNE only; cells stained red are positive only for VEGF, Ang2, Tie2, GAPDH, PKM2, GLUT1 and ATP5B. Arrows indicate examples of co-localisation. Magnification of photomicrographs × 20, insets : Figure S4

    Article Snippet: ST sections were fixed with acetone for 10 minutes and co-incubated with primary mouse antibody against human 4-HNE (GENTAUR, Kampenhout, Belgium) and with primary rabbit antibodies against VEGF, Ang2, Tie2, ATP5B and glucose transporter 1 (GLUT1) (all from Abcam), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Trevigen, Gaithersburg, MD, USA) and pyruvate kinase isozyme 2 (PKM2) (Abgent, San Diego, CA, USA).

    Techniques: Double Immunofluorescence Staining, Incubation, Marker, Immunofluorescence, Staining