l glutamine  (GE Healthcare)

 
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    Name:
    L glutamine
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    Catalog Number:
    sh30517.01
    Price:
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    5 L
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    Structured Review

    GE Healthcare l glutamine

    https://www.bioz.com/result/l glutamine/product/GE Healthcare
    Average 86 stars, based on 1 article reviews
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    Cell Culture:

    Article Title: Acute changes in liver tumour perfusion measured non-invasively with arterial spin labelling
    Article Snippet: .. Cell lines and tissue culture The human colorectal carcinoma cell lines SW1222 and LS174T ( ) were cultured in Dulbecco's Modified Eagles Medium (PAA, GE Healthcare, Fairfield, CT, USA) containing 5 mM L -glutamine (PAA, GE Healthcare) and 10% v/v Fetal bovine serum (Life Technologies, Carlsbad, CA, USA). ..

    Article Title: The Impact of Subcellular Location on the Near Infrared-Mediated Thermal Ablation of Cells by Targeted Carbon Nanotubes
    Article Snippet: Dulbecco’s modified Eagle medium (DMEM) and trypsin were purchased from Gibco (Grand Island, NY). .. The standard cell culture medium contains DMEM supplemented with 10 mM HEPES, 3.7 g/L sodium bicarbonate, 4.5 g/L D-glucose, 0.584 g/L L-glutamine, and 10% (v/v) fetal bovine serum (FBS, purchased from Hyclone, Logan, UT). .. Standard incubation conditions were at 37 °C in a humidified 10% CO2 incubator.

    Article Title: Preventative role of interleukin-17 producing regulatory T helper type 17 (Treg17) cells in type 1 diabetes in non-obese diabetic mice
    Article Snippet: .. Cells were cultured for 4 or 5 days, as stated, in complete RPMI [RPMI-1640 medium supplemented with 2 mM L-glutamine, 0.5% HEPES, 5 μg/ml penicillin, 100 U/ml streptomycin and 10% (v/v) fetal calf serum (HyClone Laboratories, Logan, UT, USA)]. .. For analysis of tissue lymphocytes by quantitative real-time reverse transcription–quantitative polymerase chain reaction (RT–qPCR), cells were extracted and restimulated as described above.

    Article Title: Urothelial bladder cancer may suppress perforin expression in CD8+ T cells by an ICAM-1/TGFβ2 mediated pathway
    Article Snippet: To explore the effects exerted by secreted factors of UBC tumor cells on CD8+ T cells, sorted CD8+ T cells from PB of healthy donors were cultured in vitro in the presence of culture supernatants from RT4 (non-muscle invasive; ATCC) and 5637 (muscle invasive; ATCC) UBC cells lines. .. Initially, both cell lines were cultured in RPMI medium (Sigma Aldrich), supplemented with 10% fetal calf serum (FCS) (Gibco), 1% L-glutamine (Hyclone), and 1% penicillin/streptomycin (Hyclone) at 37°C and 5% CO2 . .. Once the cells reached 90% confluency, the conditioned medium was removed and the cell layers were washed three times with PBS and two times with RPMI-Serum and Phenol Red Free Medium (SFM) (Thermo Fisher).

    Article Title: Treated HIV Infection Alters Phenotype But Not HIV-specific Function of Peripheral Blood Natural Killer Cells
    Article Snippet: CD4 and NK cell sorting and cell culture PBMCs were thawed, and stained with a panel consisting of 7-AAD viability staining solution (eBioscience), CD14-BV421 (clone M5E2), CD19-BV421 (clone HIB19), CD16-FITC (clone 3G8), CD3-PE (clone SK7), CD4-BV711 (clone OKT4), and CD56-PE Cy7 (clone HCD56, all antibodies from Biolegend), and sorted for CD4 T cells (CD14− CD19− CD3+ CD4+ ) and NK cells (CD14− CD19− CD3− CD56/CD16+) using a Sony SH800 sorter. .. Post-sorting, all cells were cultured in RPMI (Gibco), with 10% FBS (Thermo Fisher), 1% L-glutamine (Hyclone) and 1% penicillin/streptomycin/amphotericin (Thermo Fisher) (RP10). .. CD4 T cells were plated in RP10 with plate-bound anti-CD3 (clone OKT3, eBioscience), anti-CD28/CD49d (BD Biosciences) and PHA-L (eBioscience) for 48h.

    Article Title: The Expression of Human Cytomegalovirus MicroRNA MiR-UL148D during Latent Infection in Primary Myeloid Cells Inhibits Activin A-triggered Secretion of IL-6
    Article Snippet: The purity of isolated cells were analysed by flow cytometry, which showed 98.1% cells expressed CD14 (range 97.4–98.9%, n = 5). .. Both primary CD34+ cells and monocytes were cultured in X-Vivo-15 (Lonza) supplemented with 2.5 mM L-glutamine (GE healthcare). .. HFFF2 fibroblasts were cultured in EMEM-10, whilst KG-1 cells were maintained in RPMI-10.

    Article Title: IRF8-dependent molecular complexes control the Th9 transcriptional program
    Article Snippet: Fragments were amplified by high-fidelity PCR with C57BL/6 mice DNA as the template and specific primers (Supplementary Table ). .. Mouse NIH-3T3 cells (cultured in DMEM 4.5 g/l + glutamine + FBS at 37 °C, 5% CO2 ) were transiently transfected for 48 h with reporter plasmid and the pCMV-SPORT6-IRF8 (GE Healthcare), pCMV-SPORT6-PU.1 (GE Healthcare), pCR4-TOPO-IRF4 (Thermo Scientific) and pcDNA3.1-mBATF (Addgene) using Lipofectamine 2000 (Invitrogen). .. Luciferase was measured using the Dual Glo Luciferase Assay System (E2920, Promega) according to the manufacturer’s instructions.

    Modification:

    Article Title: Acute changes in liver tumour perfusion measured non-invasively with arterial spin labelling
    Article Snippet: .. Cell lines and tissue culture The human colorectal carcinoma cell lines SW1222 and LS174T ( ) were cultured in Dulbecco's Modified Eagles Medium (PAA, GE Healthcare, Fairfield, CT, USA) containing 5 mM L -glutamine (PAA, GE Healthcare) and 10% v/v Fetal bovine serum (Life Technologies, Carlsbad, CA, USA). ..

    Transfection:

    Article Title: IRF8-dependent molecular complexes control the Th9 transcriptional program
    Article Snippet: Fragments were amplified by high-fidelity PCR with C57BL/6 mice DNA as the template and specific primers (Supplementary Table ). .. Mouse NIH-3T3 cells (cultured in DMEM 4.5 g/l + glutamine + FBS at 37 °C, 5% CO2 ) were transiently transfected for 48 h with reporter plasmid and the pCMV-SPORT6-IRF8 (GE Healthcare), pCMV-SPORT6-PU.1 (GE Healthcare), pCR4-TOPO-IRF4 (Thermo Scientific) and pcDNA3.1-mBATF (Addgene) using Lipofectamine 2000 (Invitrogen). .. Luciferase was measured using the Dual Glo Luciferase Assay System (E2920, Promega) according to the manufacturer’s instructions.

    Plasmid Preparation:

    Article Title: IRF8-dependent molecular complexes control the Th9 transcriptional program
    Article Snippet: Fragments were amplified by high-fidelity PCR with C57BL/6 mice DNA as the template and specific primers (Supplementary Table ). .. Mouse NIH-3T3 cells (cultured in DMEM 4.5 g/l + glutamine + FBS at 37 °C, 5% CO2 ) were transiently transfected for 48 h with reporter plasmid and the pCMV-SPORT6-IRF8 (GE Healthcare), pCMV-SPORT6-PU.1 (GE Healthcare), pCR4-TOPO-IRF4 (Thermo Scientific) and pcDNA3.1-mBATF (Addgene) using Lipofectamine 2000 (Invitrogen). .. Luciferase was measured using the Dual Glo Luciferase Assay System (E2920, Promega) according to the manufacturer’s instructions.

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  • 99
    GE Healthcare mouse nih 3t3 cells
    IRF8 needs cooperative factors to induce Th9 polarisation. a Transactivation of the Il9 (left) and Il21 (right) promoters by IRF8. Il9 or Il21 promoter reporter plasmids were transfected into <t>NIH-3T3</t> cells with an IRF8-encoding vector. b Determination of IRF8-binding motifs by analysis of IRF8 ChIP-Seq peaks from WT Th9 cells. Left, letter size indicates frequency of nucleotide. Right, significance of motif occurrence. c Immunoprecipitation assay on Th9 cells differentiated for 24 h using anti-IRF8 antibody (above) or anti-PU.1 antibody (below), followed by immunoblot analysis with indicated specific antibodies. Ig is used as a control. d , e PLA in WT naive CD4 + T cells and in Th9 cells after 24 h of differentiation. DAPI staining (blue) indicates the nucleus. Green dots represent proximity between IRF8/PU.1, IRF4/PU.1 or IRF8/IRF4 ( d ) or between BATF and IRF8 or PU.1 or IRF4 ( e ) Magnifications are ×63, scale bar represents 5 µm. f Top panel represent a screenshot from the ECR (evolutionary conserved regions) Browser web site of the mouse Il9 (left) and Il21 (right) genes. Exons are in blue, introns in pink, UTRs in yellow and CNS are in red. Bottom panel is a representation of binding peaks of ChIP-Seq data of BATF, IRF4, PU.1 and IRF8 in WT Th9 cells differentiated for 24 h. The red square shows the overlap between BATF, IRF4, PU.1 and IRF8 peaks in Il9 or Il21 CNS1. g Co-location of IRF8, PU.1, IRF4 and BATF-binding peaks in Th9 cells were determined by multiple overlap analysis of the respective ChIP-Seq data. IRF8 can bind DNA alone or in combination with one, two or three factors (IRF4, BATF and PU.1). h The average density of IRF4, PU.1 and BATF in Th9 cells is shown for regions centred on the summits of the IRF8 peaks. i Transactivation of the Il9 promoters by IRF8, PU.1, IRF4 and BATF. Il9 promoter reporter plasmids were transfected into NIH-3T3 cells with vectors encoding IRF8, PU.1, IRF4 and BATF alone or in combination. ns, not significant; * P
    Mouse Nih 3t3 Cells, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GE Healthcare phenol red free rpmi 1640
    Depletion of androgen increased AR expression and Akt activity LNCaP cells were grown in regular <t>RPMI</t> 1640 medium with 10% FBS and androgen-depleted medium, phenol red-free RPMI 1640 supplemented with 10% charcoal/dextran-treated FBS for 10 days. Cells were lyzed and 50 μg total protein was resolved by electrophoresis on a 10% SDS-PAGE gel. Immunoblotting was performed using AR and phospho-Akt antibodies, respectively. β-actin protein was used as a loading control.
    Phenol Red Free Rpmi 1640, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    GE Healthcare dmem high glucose
    MGF inhibits lactic acidosis. A : ECAR recorded in the same Seahorse Bioscience Metabolic Flux experiments as shown in Fig. 4 E . B : Lactate formed in the experiments shown in A was measured by a d -lactate assay kit. C : <t>C2C12</t> myotubes were incubated overnight in glucose-free and sodium pyruvate–free <t>DMEM</t> containing l -glutamine and 2% horse serum, in the presence of various concentrations of MGF, and then incubated in media containing 25 mmol/L glucose in the presence of matching concentrations of MGF for 8 h. Media were collected, and lactate in the media was measured by a d -lactate assay kit (Eton Biosciences). These experiments were all conducted three to four times and each time with triplicates for each condition. Values are average ± SEM. ** P
    Dmem High Glucose, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    IRF8 needs cooperative factors to induce Th9 polarisation. a Transactivation of the Il9 (left) and Il21 (right) promoters by IRF8. Il9 or Il21 promoter reporter plasmids were transfected into NIH-3T3 cells with an IRF8-encoding vector. b Determination of IRF8-binding motifs by analysis of IRF8 ChIP-Seq peaks from WT Th9 cells. Left, letter size indicates frequency of nucleotide. Right, significance of motif occurrence. c Immunoprecipitation assay on Th9 cells differentiated for 24 h using anti-IRF8 antibody (above) or anti-PU.1 antibody (below), followed by immunoblot analysis with indicated specific antibodies. Ig is used as a control. d , e PLA in WT naive CD4 + T cells and in Th9 cells after 24 h of differentiation. DAPI staining (blue) indicates the nucleus. Green dots represent proximity between IRF8/PU.1, IRF4/PU.1 or IRF8/IRF4 ( d ) or between BATF and IRF8 or PU.1 or IRF4 ( e ) Magnifications are ×63, scale bar represents 5 µm. f Top panel represent a screenshot from the ECR (evolutionary conserved regions) Browser web site of the mouse Il9 (left) and Il21 (right) genes. Exons are in blue, introns in pink, UTRs in yellow and CNS are in red. Bottom panel is a representation of binding peaks of ChIP-Seq data of BATF, IRF4, PU.1 and IRF8 in WT Th9 cells differentiated for 24 h. The red square shows the overlap between BATF, IRF4, PU.1 and IRF8 peaks in Il9 or Il21 CNS1. g Co-location of IRF8, PU.1, IRF4 and BATF-binding peaks in Th9 cells were determined by multiple overlap analysis of the respective ChIP-Seq data. IRF8 can bind DNA alone or in combination with one, two or three factors (IRF4, BATF and PU.1). h The average density of IRF4, PU.1 and BATF in Th9 cells is shown for regions centred on the summits of the IRF8 peaks. i Transactivation of the Il9 promoters by IRF8, PU.1, IRF4 and BATF. Il9 promoter reporter plasmids were transfected into NIH-3T3 cells with vectors encoding IRF8, PU.1, IRF4 and BATF alone or in combination. ns, not significant; * P

    Journal: Nature Communications

    Article Title: IRF8-dependent molecular complexes control the Th9 transcriptional program

    doi: 10.1038/s41467-017-01070-w

    Figure Lengend Snippet: IRF8 needs cooperative factors to induce Th9 polarisation. a Transactivation of the Il9 (left) and Il21 (right) promoters by IRF8. Il9 or Il21 promoter reporter plasmids were transfected into NIH-3T3 cells with an IRF8-encoding vector. b Determination of IRF8-binding motifs by analysis of IRF8 ChIP-Seq peaks from WT Th9 cells. Left, letter size indicates frequency of nucleotide. Right, significance of motif occurrence. c Immunoprecipitation assay on Th9 cells differentiated for 24 h using anti-IRF8 antibody (above) or anti-PU.1 antibody (below), followed by immunoblot analysis with indicated specific antibodies. Ig is used as a control. d , e PLA in WT naive CD4 + T cells and in Th9 cells after 24 h of differentiation. DAPI staining (blue) indicates the nucleus. Green dots represent proximity between IRF8/PU.1, IRF4/PU.1 or IRF8/IRF4 ( d ) or between BATF and IRF8 or PU.1 or IRF4 ( e ) Magnifications are ×63, scale bar represents 5 µm. f Top panel represent a screenshot from the ECR (evolutionary conserved regions) Browser web site of the mouse Il9 (left) and Il21 (right) genes. Exons are in blue, introns in pink, UTRs in yellow and CNS are in red. Bottom panel is a representation of binding peaks of ChIP-Seq data of BATF, IRF4, PU.1 and IRF8 in WT Th9 cells differentiated for 24 h. The red square shows the overlap between BATF, IRF4, PU.1 and IRF8 peaks in Il9 or Il21 CNS1. g Co-location of IRF8, PU.1, IRF4 and BATF-binding peaks in Th9 cells were determined by multiple overlap analysis of the respective ChIP-Seq data. IRF8 can bind DNA alone or in combination with one, two or three factors (IRF4, BATF and PU.1). h The average density of IRF4, PU.1 and BATF in Th9 cells is shown for regions centred on the summits of the IRF8 peaks. i Transactivation of the Il9 promoters by IRF8, PU.1, IRF4 and BATF. Il9 promoter reporter plasmids were transfected into NIH-3T3 cells with vectors encoding IRF8, PU.1, IRF4 and BATF alone or in combination. ns, not significant; * P

    Article Snippet: Mouse NIH-3T3 cells (cultured in DMEM 4.5 g/l + glutamine + FBS at 37 °C, 5% CO2 ) were transiently transfected for 48 h with reporter plasmid and the pCMV-SPORT6-IRF8 (GE Healthcare), pCMV-SPORT6-PU.1 (GE Healthcare), pCR4-TOPO-IRF4 (Thermo Scientific) and pcDNA3.1-mBATF (Addgene) using Lipofectamine 2000 (Invitrogen).

    Techniques: Transfection, Plasmid Preparation, Binding Assay, Chromatin Immunoprecipitation, Immunoprecipitation, Proximity Ligation Assay, Staining

    Depletion of androgen increased AR expression and Akt activity LNCaP cells were grown in regular RPMI 1640 medium with 10% FBS and androgen-depleted medium, phenol red-free RPMI 1640 supplemented with 10% charcoal/dextran-treated FBS for 10 days. Cells were lyzed and 50 μg total protein was resolved by electrophoresis on a 10% SDS-PAGE gel. Immunoblotting was performed using AR and phospho-Akt antibodies, respectively. β-actin protein was used as a loading control.

    Journal: Oncotarget

    Article Title: Reciprocal feedback inhibition of the androgen receptor and PI3K as a novel therapy for castrate-sensitive and -resistant prostate cancer

    doi:

    Figure Lengend Snippet: Depletion of androgen increased AR expression and Akt activity LNCaP cells were grown in regular RPMI 1640 medium with 10% FBS and androgen-depleted medium, phenol red-free RPMI 1640 supplemented with 10% charcoal/dextran-treated FBS for 10 days. Cells were lyzed and 50 μg total protein was resolved by electrophoresis on a 10% SDS-PAGE gel. Immunoblotting was performed using AR and phospho-Akt antibodies, respectively. β-actin protein was used as a loading control.

    Article Snippet: For the androgen-depletion experiments, LNCaP cells were grown in androgen-depleted medium, phenol red-free RPMI 1640 supplemented with 10% charcoal/dextran-treated FBS (HyClone, Logan, UT).

    Techniques: Expressing, Activity Assay, Electrophoresis, SDS Page

    MGF inhibits lactic acidosis. A : ECAR recorded in the same Seahorse Bioscience Metabolic Flux experiments as shown in Fig. 4 E . B : Lactate formed in the experiments shown in A was measured by a d -lactate assay kit. C : C2C12 myotubes were incubated overnight in glucose-free and sodium pyruvate–free DMEM containing l -glutamine and 2% horse serum, in the presence of various concentrations of MGF, and then incubated in media containing 25 mmol/L glucose in the presence of matching concentrations of MGF for 8 h. Media were collected, and lactate in the media was measured by a d -lactate assay kit (Eton Biosciences). These experiments were all conducted three to four times and each time with triplicates for each condition. Values are average ± SEM. ** P

    Journal: Diabetes

    Article Title: Mangiferin Stimulates Carbohydrate Oxidation and Protects Against Metabolic Disorders Induced by High-Fat Diets

    doi: 10.2337/db14-0006

    Figure Lengend Snippet: MGF inhibits lactic acidosis. A : ECAR recorded in the same Seahorse Bioscience Metabolic Flux experiments as shown in Fig. 4 E . B : Lactate formed in the experiments shown in A was measured by a d -lactate assay kit. C : C2C12 myotubes were incubated overnight in glucose-free and sodium pyruvate–free DMEM containing l -glutamine and 2% horse serum, in the presence of various concentrations of MGF, and then incubated in media containing 25 mmol/L glucose in the presence of matching concentrations of MGF for 8 h. Media were collected, and lactate in the media was measured by a d -lactate assay kit (Eton Biosciences). These experiments were all conducted three to four times and each time with triplicates for each condition. Values are average ± SEM. ** P

    Article Snippet: Cell Culture C2C12 myoblast cells were maintained in DMEM high glucose (HyClone Laboratories, Logan, UT), supplemented with 1% penicillin and streptomycin (Gibco, Carlsbad, CA) and 10% FBS.

    Techniques: Lactate Assay, Incubation

    MGF stimulates glucose oxidation. Palmitate ( A ) and glucose ( B ) oxidation was measured after a 12-h fast in soleus muscle isolated from mice fed with CD or HFD, with or without 0.5% MGF, for 18 weeks ( n = 6–7). C : [ 14 C]-labeled oleic acid oxidation in C2C12 myotubes pretreated or not with 200 μmol/L MGF for 16 h. Palmitate ( D ) and glucose ( E ) oxidation in C2C12 myotubes measured by Seahorse Bioscience Metabolic Flux analyzer. Myotubes differentiated from C2C12 cells in XF24 microplates were incubated overnight in glucose-free and sodium pyruvate–free DMEM, with or without 200 μmol/L MGF. OCR was recorded after injection of vehicle ( D ), BSA-conjugated palmitate ( D ), or d -glucose ( E ) into wells, with or without 200 μmol/L MGF. F : Intracellular ATP levels in C2C12 myotubes incubated overnight in glucose-free and sodium pyruvate–free DMEM in the presence of various concentrations of MGF and then incubated in media containing 25 mmol/L glucose with matching concentrations of MGF for 8 h. ATP was determined using the CellTiter-Glo Luminescent Cell Viability Assay kit according to the manufacturer’s instructions (Promega). In vitro experiments were conducted three to four times and each time with triplicates for each condition. Values are average ± SEM. * P

    Journal: Diabetes

    Article Title: Mangiferin Stimulates Carbohydrate Oxidation and Protects Against Metabolic Disorders Induced by High-Fat Diets

    doi: 10.2337/db14-0006

    Figure Lengend Snippet: MGF stimulates glucose oxidation. Palmitate ( A ) and glucose ( B ) oxidation was measured after a 12-h fast in soleus muscle isolated from mice fed with CD or HFD, with or without 0.5% MGF, for 18 weeks ( n = 6–7). C : [ 14 C]-labeled oleic acid oxidation in C2C12 myotubes pretreated or not with 200 μmol/L MGF for 16 h. Palmitate ( D ) and glucose ( E ) oxidation in C2C12 myotubes measured by Seahorse Bioscience Metabolic Flux analyzer. Myotubes differentiated from C2C12 cells in XF24 microplates were incubated overnight in glucose-free and sodium pyruvate–free DMEM, with or without 200 μmol/L MGF. OCR was recorded after injection of vehicle ( D ), BSA-conjugated palmitate ( D ), or d -glucose ( E ) into wells, with or without 200 μmol/L MGF. F : Intracellular ATP levels in C2C12 myotubes incubated overnight in glucose-free and sodium pyruvate–free DMEM in the presence of various concentrations of MGF and then incubated in media containing 25 mmol/L glucose with matching concentrations of MGF for 8 h. ATP was determined using the CellTiter-Glo Luminescent Cell Viability Assay kit according to the manufacturer’s instructions (Promega). In vitro experiments were conducted three to four times and each time with triplicates for each condition. Values are average ± SEM. * P

    Article Snippet: Cell Culture C2C12 myoblast cells were maintained in DMEM high glucose (HyClone Laboratories, Logan, UT), supplemented with 1% penicillin and streptomycin (Gibco, Carlsbad, CA) and 10% FBS.

    Techniques: Isolation, Mouse Assay, Labeling, Incubation, Injection, Cell Viability Assay, In Vitro

    MGF enhances pyruvate oxidation by activating PDH. A : Pyruvate oxidation in myotubes differentiated from C2C12 cells measured by Seahorse Bioscience Metabolic Flux analysis. Myotubes differentiated from C2C12 cells in XF24 microplates were incubated overnight in glucose-free and sodium pyruvate–free DMEM, with and without 200 μmol/L MGF. OCR was recorded after injection of sodium pyruvate into wells, with or without 200 μmol/L MGF. B : PDH activities measured as the difference between absorbances at 500 nm and 750 nm at 25°C in mitochondrial protein from C2C12 myotubes treated with various concentrations of MGF for 24 h or 15 min or with 5 mmol/L DCA or FP + 200 μmol/L MGF for 15 min. C : Initial rates of PDH-catalyzed pyruvate oxidation measured as the difference between absorbances at 500 nm and 750 nm in mitochondrial protein from C2C12 myotubes treated with various concentrations of MGF for 24 h or 15 min. The unit of activity was determined by using extinction coefficient of reduced p-iodonitrotetrazolium violet (12.4 mmol ⋅ L −1 ⋅ cm −1 ). D : Initial rates of PDH catalyzed [ 14 C]-labeled pyruvate oxidation measured as the production of 14 CO 2 by mitochondrial protein from C2C12 myotubes treated with various concentrations of MGF for 15 min. E : Western blots of proteins extracted from C2C12 myotubes treated with 200 or 1,000 μmol/L MGF for 15 min and 24 h. Membrane was blotted with PDH (1:250 dilution; Cell Signaling), p-PDH (1:2,000; Abcam), PDK4 (1:500; laboratory of Dr. Robert Harris at Indiana University School of Medicine), or α-tubulin (1:2,000; Sigma-Aldrich) at 4°C overnight. F : Quantitative analysis of Western blots shown in E . G : Western blots of proteins extracted from C2C12 myotubes treated with 200 or 1,000 μmol/L MGF or 5 mmol/L DCA for 15 min. H : Quantitative analysis of Western blots shown in G . Values are average ± SEM. * P

    Journal: Diabetes

    Article Title: Mangiferin Stimulates Carbohydrate Oxidation and Protects Against Metabolic Disorders Induced by High-Fat Diets

    doi: 10.2337/db14-0006

    Figure Lengend Snippet: MGF enhances pyruvate oxidation by activating PDH. A : Pyruvate oxidation in myotubes differentiated from C2C12 cells measured by Seahorse Bioscience Metabolic Flux analysis. Myotubes differentiated from C2C12 cells in XF24 microplates were incubated overnight in glucose-free and sodium pyruvate–free DMEM, with and without 200 μmol/L MGF. OCR was recorded after injection of sodium pyruvate into wells, with or without 200 μmol/L MGF. B : PDH activities measured as the difference between absorbances at 500 nm and 750 nm at 25°C in mitochondrial protein from C2C12 myotubes treated with various concentrations of MGF for 24 h or 15 min or with 5 mmol/L DCA or FP + 200 μmol/L MGF for 15 min. C : Initial rates of PDH-catalyzed pyruvate oxidation measured as the difference between absorbances at 500 nm and 750 nm in mitochondrial protein from C2C12 myotubes treated with various concentrations of MGF for 24 h or 15 min. The unit of activity was determined by using extinction coefficient of reduced p-iodonitrotetrazolium violet (12.4 mmol ⋅ L −1 ⋅ cm −1 ). D : Initial rates of PDH catalyzed [ 14 C]-labeled pyruvate oxidation measured as the production of 14 CO 2 by mitochondrial protein from C2C12 myotubes treated with various concentrations of MGF for 15 min. E : Western blots of proteins extracted from C2C12 myotubes treated with 200 or 1,000 μmol/L MGF for 15 min and 24 h. Membrane was blotted with PDH (1:250 dilution; Cell Signaling), p-PDH (1:2,000; Abcam), PDK4 (1:500; laboratory of Dr. Robert Harris at Indiana University School of Medicine), or α-tubulin (1:2,000; Sigma-Aldrich) at 4°C overnight. F : Quantitative analysis of Western blots shown in E . G : Western blots of proteins extracted from C2C12 myotubes treated with 200 or 1,000 μmol/L MGF or 5 mmol/L DCA for 15 min. H : Quantitative analysis of Western blots shown in G . Values are average ± SEM. * P

    Article Snippet: Cell Culture C2C12 myoblast cells were maintained in DMEM high glucose (HyClone Laboratories, Logan, UT), supplemented with 1% penicillin and streptomycin (Gibco, Carlsbad, CA) and 10% FBS.

    Techniques: Incubation, Injection, Activity Assay, Labeling, Western Blot

    Effects of Thd on HG-induced MC viability by MTT assay. NG-cultured MCs were grown in DMEM containing 5.6 mmol/l D-glucose, whilst HG-cultured MCs were grown in Dulbecco's modified Eagle's medium containing 30 mmol/l D-glucose and following treatment with various concentrations (0, 10, 50, 100 and 200 µg/ml) of Thd for 12, 24 or 48 h the MTT assay was used to measure cell viability. Data are presented as the mean ± standard error of the mean (n=5). # P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Thalidomide decreases high glucose-induced extracellular matrix protein synthesis in mesangial cells via the AMPK pathway

    doi: 10.3892/etm.2018.6995

    Figure Lengend Snippet: Effects of Thd on HG-induced MC viability by MTT assay. NG-cultured MCs were grown in DMEM containing 5.6 mmol/l D-glucose, whilst HG-cultured MCs were grown in Dulbecco's modified Eagle's medium containing 30 mmol/l D-glucose and following treatment with various concentrations (0, 10, 50, 100 and 200 µg/ml) of Thd for 12, 24 or 48 h the MTT assay was used to measure cell viability. Data are presented as the mean ± standard error of the mean (n=5). # P

    Article Snippet: Cells were cultured in Dulbecco's modified Eagle's media (DMEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; HyClone; GE Healthcare Life Sciences) and maintained at 37°C in a 5% CO2 -humidified incubator.

    Techniques: MTT Assay, Cell Culture, Modification