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  • 97
    Name:
    Gas Chromatography
    Description:
    This book presents an introduction to gas chromatography including examples of applications Shows readers malfunctions that can occur enabling them to recognize and fix faults
    Catalog Number:
    z271756
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    None
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    Structured Review

    Millipore gas chromatograph
    Gas Chromatography
    This book presents an introduction to gas chromatography including examples of applications Shows readers malfunctions that can occur enabling them to recognize and fix faults
    https://www.bioz.com/result/gas chromatograph/product/Millipore
    Average 97 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    gas chromatograph - by Bioz Stars, 2020-07
    97/100 stars

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    Related Articles

    Gas Chromatography:

    Article Title: Toward aldehyde and alkane production by removing aldehyde reductase activity in Escherichia coli
    Article Snippet: .. All chemicals for gas chromatography (GC) standards except for ethanol and 1-pentanol were purchased from Sigma-Aldrich (St. Louis, MO). .. 1-Pentanol was purchased from Acros Organics (Belgium).

    Article Title: Crystallisation properties of amorphous cyclodextrin powders and their complexation with fish oil
    Article Snippet: .. Absolute ethanol, gas chromatography grade hexane and fish oil (derived from the Menhaden fish.) were purchased from Sigma–Aldrich (Sigma–Aldrich Co. LLC, Australia). ..

    Article Title: Combined Use of S. pombe and L. thermotolerans in Winemaking. Beneficial Effects Determined through the Study of Wines’ Analytical Characteristics
    Article Snippet: .. We used gas chromatography quality compounds as the sets of the volatile standards for this purpose (Fluka, Sigma-Aldrich Corp., Buchs SG, Standort, Switzerland). ..

    Article Title: Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide
    Article Snippet: .. Determination of inhibitory activity of the lysate of m3MST-overexpressing cells by means of gas chromatography HEK 293 cells in ice-cold isolation buffer containing 30 mM HEPES buffer (pH 7.4), 100 μM DTT and 1% protease inhibitor cocktail (Sigma, Japan) were sonicated for 10 sec twice in an Astrason 4000 sonicator (MISONIX, USA) to obtain the lysate. ..

    Chromatography:

    Article Title: The genome and phenome of the green alga Chloroidium sp. UTEX 3007 reveal adaptive traits for desert acclimatization
    Article Snippet: .. Gas chromatography-flame ionization detection (GC-FID) For the extraction, 50 mg algal material (dry weight) was used by adding 1 ml of 1 N HCl/methanol solution (Sigma-Aldrich, Darmstadt, Germany). .. An internal standard (100 µl of FA15:0, pentadecanoic acid) was added to each sample before incubation at 80°C in a water bath for 30 min. After cooling to room temperature, 1 ml of 0.9% NaCl and 1 ml of 100% hexane were added to each vial.

    Isolation:

    Article Title: Mitochondrial protein functions elucidated by multi-omic mass spectrometry profiling
    Article Snippet: .. Gas chromatography-mass spectrometry (GC-MS) metabolomics 1×108 yeast cells yeast cells were isolated by rapid vacuum filtration onto a nylon filter membrane (0.45 μm pore size, Millipore) using a Glass Microanalysis Filter Holder (Millipore), briefly washed with phosphate buffered saline (1 mL), and immediately submerged into ACN/MeOH/H2 O (2:2:1, v/v/v, 1.5 mL, pre-cooled to −20 °C) in a plastic tube. .. The time from sampling yeast from the culture to submersion in cold extraction solvent was less than 30 s. Tubes with the extraction solvent, nylon filter, and yeast were stored at −80 °C prior to analysis.

    Article Title: Rapid assessment of the authenticity of limequat fruit using the electronic nose and gas chromatography coupled with mass spectrometry
    Article Snippet: .. Gas chromatography–mass spectrometry analysis Solid phase microextraction was utilized for the isolation and enrichment of analytes using divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS)-coated fibre with thickness of 50/30 µm and length of 2 cm (Sigma-Aldrich). .. Extraction was done at 40 °C for 35 min. Next, the SPME fibre with extracted analytes was automatically transferred into the injector port of the gas chromatograph for thermal desorption for 6 min.

    Article Title: Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide
    Article Snippet: .. Determination of inhibitory activity of the lysate of m3MST-overexpressing cells by means of gas chromatography HEK 293 cells in ice-cold isolation buffer containing 30 mM HEPES buffer (pH 7.4), 100 μM DTT and 1% protease inhibitor cocktail (Sigma, Japan) were sonicated for 10 sec twice in an Astrason 4000 sonicator (MISONIX, USA) to obtain the lysate. ..

    Solid-phase Microextraction:

    Article Title: Rapid assessment of the authenticity of limequat fruit using the electronic nose and gas chromatography coupled with mass spectrometry
    Article Snippet: .. Gas chromatography–mass spectrometry analysis Solid phase microextraction was utilized for the isolation and enrichment of analytes using divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS)-coated fibre with thickness of 50/30 µm and length of 2 cm (Sigma-Aldrich). .. Extraction was done at 40 °C for 35 min. Next, the SPME fibre with extracted analytes was automatically transferred into the injector port of the gas chromatograph for thermal desorption for 6 min.

    Size-exclusion Chromatography:

    Article Title: Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide
    Article Snippet: .. Determination of inhibitory activity of the lysate of m3MST-overexpressing cells by means of gas chromatography HEK 293 cells in ice-cold isolation buffer containing 30 mM HEPES buffer (pH 7.4), 100 μM DTT and 1% protease inhibitor cocktail (Sigma, Japan) were sonicated for 10 sec twice in an Astrason 4000 sonicator (MISONIX, USA) to obtain the lysate. ..

    Suction Filtration:

    Article Title: Mitochondrial protein functions elucidated by multi-omic mass spectrometry profiling
    Article Snippet: .. Gas chromatography-mass spectrometry (GC-MS) metabolomics 1×108 yeast cells yeast cells were isolated by rapid vacuum filtration onto a nylon filter membrane (0.45 μm pore size, Millipore) using a Glass Microanalysis Filter Holder (Millipore), briefly washed with phosphate buffered saline (1 mL), and immediately submerged into ACN/MeOH/H2 O (2:2:1, v/v/v, 1.5 mL, pre-cooled to −20 °C) in a plastic tube. .. The time from sampling yeast from the culture to submersion in cold extraction solvent was less than 30 s. Tubes with the extraction solvent, nylon filter, and yeast were stored at −80 °C prior to analysis.

    Gas Chromatography-Mass Spectrometry:

    Article Title: Mitochondrial protein functions elucidated by multi-omic mass spectrometry profiling
    Article Snippet: .. Gas chromatography-mass spectrometry (GC-MS) metabolomics 1×108 yeast cells yeast cells were isolated by rapid vacuum filtration onto a nylon filter membrane (0.45 μm pore size, Millipore) using a Glass Microanalysis Filter Holder (Millipore), briefly washed with phosphate buffered saline (1 mL), and immediately submerged into ACN/MeOH/H2 O (2:2:1, v/v/v, 1.5 mL, pre-cooled to −20 °C) in a plastic tube. .. The time from sampling yeast from the culture to submersion in cold extraction solvent was less than 30 s. Tubes with the extraction solvent, nylon filter, and yeast were stored at −80 °C prior to analysis.

    Article Title: Rapid assessment of the authenticity of limequat fruit using the electronic nose and gas chromatography coupled with mass spectrometry
    Article Snippet: .. Gas chromatography–mass spectrometry analysis Solid phase microextraction was utilized for the isolation and enrichment of analytes using divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS)-coated fibre with thickness of 50/30 µm and length of 2 cm (Sigma-Aldrich). .. Extraction was done at 40 °C for 35 min. Next, the SPME fibre with extracted analytes was automatically transferred into the injector port of the gas chromatograph for thermal desorption for 6 min.

    Article Title: Caenorhabditis elegans Battling Starvation Stress: Low Levels of Ethanol Prolong Lifespan in L1 Larvae
    Article Snippet: .. Fatty acid analysis by gas chromatography-mass spectrometry (GC-MS) L1 larvae prepared as described above for the survival experiments were incubated for various times at 20°C at 160 rpm in M9 medium alone, M9 medium with 1 mM ethanol, or M9 medium with 1 mM fully-deuterated ethanol (Sigma, catalog no. 186414). ..

    Incubation:

    Article Title: Caenorhabditis elegans Battling Starvation Stress: Low Levels of Ethanol Prolong Lifespan in L1 Larvae
    Article Snippet: .. Fatty acid analysis by gas chromatography-mass spectrometry (GC-MS) L1 larvae prepared as described above for the survival experiments were incubated for various times at 20°C at 160 rpm in M9 medium alone, M9 medium with 1 mM ethanol, or M9 medium with 1 mM fully-deuterated ethanol (Sigma, catalog no. 186414). ..

    Activity Assay:

    Article Title: Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide
    Article Snippet: .. Determination of inhibitory activity of the lysate of m3MST-overexpressing cells by means of gas chromatography HEK 293 cells in ice-cold isolation buffer containing 30 mM HEPES buffer (pH 7.4), 100 μM DTT and 1% protease inhibitor cocktail (Sigma, Japan) were sonicated for 10 sec twice in an Astrason 4000 sonicator (MISONIX, USA) to obtain the lysate. ..

    Protease Inhibitor:

    Article Title: Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide
    Article Snippet: .. Determination of inhibitory activity of the lysate of m3MST-overexpressing cells by means of gas chromatography HEK 293 cells in ice-cold isolation buffer containing 30 mM HEPES buffer (pH 7.4), 100 μM DTT and 1% protease inhibitor cocktail (Sigma, Japan) were sonicated for 10 sec twice in an Astrason 4000 sonicator (MISONIX, USA) to obtain the lysate. ..

    Sonication:

    Article Title: Discovery and Mechanistic Characterization of Selective Inhibitors of H2S-producing Enzyme: 3-Mercaptopyruvate Sulfurtransferase (3MST) Targeting Active-site Cysteine Persulfide
    Article Snippet: .. Determination of inhibitory activity of the lysate of m3MST-overexpressing cells by means of gas chromatography HEK 293 cells in ice-cold isolation buffer containing 30 mM HEPES buffer (pH 7.4), 100 μM DTT and 1% protease inhibitor cocktail (Sigma, Japan) were sonicated for 10 sec twice in an Astrason 4000 sonicator (MISONIX, USA) to obtain the lysate. ..

    Fluorescence In Situ Hybridization:

    Article Title: Crystallisation properties of amorphous cyclodextrin powders and their complexation with fish oil
    Article Snippet: .. Absolute ethanol, gas chromatography grade hexane and fish oil (derived from the Menhaden fish.) were purchased from Sigma–Aldrich (Sigma–Aldrich Co. LLC, Australia). ..

    Derivative Assay:

    Article Title: Crystallisation properties of amorphous cyclodextrin powders and their complexation with fish oil
    Article Snippet: .. Absolute ethanol, gas chromatography grade hexane and fish oil (derived from the Menhaden fish.) were purchased from Sigma–Aldrich (Sigma–Aldrich Co. LLC, Australia). ..

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  • 88
    Millipore p foxo
    Exercise training reduces IL-6 induced alterations in mitochondria dynamics, apoptosis and <t>FoxO</t> phosphorylation. A) Representative Western blot of Mfn1, Mfn2 and <t>FIS1</t> protein in the gastrocnemius of Apc Min/+ mice. B) Mfn1, C) Mfn2 and D) FIS1 protein expression normalized to sedentary mice treated with the control vector. E) upper - Representative Western blot of total and phosphorylated FoxO proteins; lower - ratio of phosphorylated to total FoxO protein expression normalized to cage control mice. F) Bax mRNA expression. Data are normalized to cage control mice. Values are means ± SE. Significance was set at P
    P Foxo, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p foxo/product/Millipore
    Average 88 stars, based on 3 article reviews
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    91
    Millipore ucp1 promoter
    Knockdown of Irx3 repressed the differentiation of SVFs toward beige adipocytes in vitro. (A–B) (A) Relative mRNA and (B) protein levels of Irx3 in SVFs from IWAT were efficiently knocked down after a 3-day infection of mouse Irx3 lentiviral shRNA ( n = 3 for each group). (C–D) (C) Oil Red O staining and (D) TG concentration of the mature beige adipocytes under eight-day differentiation ( n = 6 for each group). (E) Relative mRNA expression of <t>Ucp1</t> , Cidea , Pgc-1α , C/ebpβ , and Prdm16 at different time points during beige adipocyte differentiation from IWAT SVFs ( n = 3–4 for different groups). (F–G) (F) Relative mRNA ( n = 4) and (G) protein expression of brown adipocyte marker genes of the induced beige adipocytes from mouse IWAT SVTs for eight days with the infection of mouse Irx3 lentiviral shRNA, with or without CL-316,243 activation. (H) Relative mRNA expression of adipocyte differentiation-related genes in beige adipocytes ( n = 4). (I) Relative mRNA expression of the indicated genes in the induced brown adipocytes from mouse BAT SVFs for eight days, with the infection of mouse Irx3 lentiviral shRNA ( n = 4). (J) Relative mRNA levels of IRX3 in SVFs from human sWAT after a 3-day infection of human IRX3 lentiviral shRNA (left), and the mRNA levels of brown adipocyte marker genes in the SVFs under 2-day differentiation ( n = 4). (K) OCR measurement of the beige adipocytes under 5-day differentiation, with the infection of mouse Irx3 lentiviral shRNA ( n = 5). (L–N) (L) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector or IRX3 for 48 h. (M) Schematic diagram of the mutant mouse Ucp1 promoter deleting ACATGTGT among the − 3470 to − 3463 bp region proximal to the TSS of the Ucp1 gene. (N) Transcriptional activity of wild-type or mutant Ucp1 promoter was measured with IRX3 overexpression. (O) qPCR of Ucp1 promoter binding region which was recruited by Irx3 antibody in 8-day induced beige adipocytes, as analyzed by ChIP. Fold enrichment of Ucp1 promoter was given ( n = 3). For the measurement of luciferase activity, cells were seeded on 24-well plate and transfected with 800 ng pEGFP-C1 vector or IRX3, 200 ng Ucp1 promoter construct, and 1 ng pRL-SV40, followed by the harvest for luciferase activity assessment using a dual-luciferase reporter assay system (Promega). Luciferase activity was corrected for Renilla luciferase activity ( n = 3–4 for each group). For qPCR data, mRNA expression was normalized to 36b4 for mouse genes and βACTIN for human genes. SVF cells were isolated from IWAT and BAT of C57BL/6J mice or human sWAT wherever mentioned, and IRX3 shRNA were introduced to cells 24 h after seeding. Data were presented as mean ± s.e.m. * P
    Ucp1 Promoter, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore pgc 1α
    Quercetin promoted translocation of <t>PGC‐1α</t> from cytoplasm to nucleus and enhanced PGC‐1α binding. ( A‐D ) The representative photomicrographs showing PGC‐1α immunohistochemistry of tissue from different group 24 hrs after TBI. ( E, H ) The nuclear PGC‐1α expression after quercetin treatment in mice with TBI, as measured by Western blot. ( F, I ) The cytoplasmic protein PGC‐1α expression after quercetin treatment in mice with TBI, as measured by Western blot. ( G, J ) The total protein PGC‐1α expression after melatonin treatment in mice with TBI, as measured by Western blot. Bars represent the mean ± S.E.M. * P
    Pgc 1α, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 177 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore trb3
    MT prevents tBHP inhibition of Akt function in vitro. Adult cardiomyocytes from WT and MT-TG mice were directly exposed to tBHP (100 μmol/L) for 2 h or LPS (10 µg/mL) for 4 h and then stimulated with or without insulin (INS; 100 nmol/L) for 15 min. Cells were collected, and the phosphorylation of total Akt ( A and B ), Akt2 ( A , C , I , and J ), GSK-3β ( A , D , I , and K ), and GS ( A , E , I , and L ) and expression of HK II ( A , F , I , and M ), PGC-1α ( A , G , I , and N ), and <t>TRB3</t> ( A , H , I , and O ) was detected by Western blot. Results were normalized to WT control. Three independent experiments were performed. Data shown in graphs represent the means ± SD. * P
    Trb3, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Exercise training reduces IL-6 induced alterations in mitochondria dynamics, apoptosis and FoxO phosphorylation. A) Representative Western blot of Mfn1, Mfn2 and FIS1 protein in the gastrocnemius of Apc Min/+ mice. B) Mfn1, C) Mfn2 and D) FIS1 protein expression normalized to sedentary mice treated with the control vector. E) upper - Representative Western blot of total and phosphorylated FoxO proteins; lower - ratio of phosphorylated to total FoxO protein expression normalized to cage control mice. F) Bax mRNA expression. Data are normalized to cage control mice. Values are means ± SE. Significance was set at P

    Journal: Skeletal Muscle

    Article Title: IL-6 regulation on skeletal muscle mitochondrial remodeling during cancer cachexia in the ApcMin/+ mouse

    doi: 10.1186/2044-5040-2-14

    Figure Lengend Snippet: Exercise training reduces IL-6 induced alterations in mitochondria dynamics, apoptosis and FoxO phosphorylation. A) Representative Western blot of Mfn1, Mfn2 and FIS1 protein in the gastrocnemius of Apc Min/+ mice. B) Mfn1, C) Mfn2 and D) FIS1 protein expression normalized to sedentary mice treated with the control vector. E) upper - Representative Western blot of total and phosphorylated FoxO proteins; lower - ratio of phosphorylated to total FoxO protein expression normalized to cage control mice. F) Bax mRNA expression. Data are normalized to cage control mice. Values are means ± SE. Significance was set at P

    Article Snippet: Primary antibodies for CoxIV, Cytochrome C, Atg5, Beclin-1, LC3β, GAPDH and FoxO (Cell Signaling, Danvers, MA, USA), Mfn1, Mfn2 (Novus Biologicals, Littleton, CO, USA), Fis1 (Sigma), PGC-1α (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-FoxO (Millipore, Billerica, MA, USA) and 4-hydroxynonenal (alpha diagnostics) were diluted 1:1,000 to 1:500 in 5% milk in TBS-T followed by 1 h incubation with membranes at room temperature.

    Techniques: Western Blot, Mouse Assay, Expressing, Plasmid Preparation

    Knockdown of Irx3 repressed the differentiation of SVFs toward beige adipocytes in vitro. (A–B) (A) Relative mRNA and (B) protein levels of Irx3 in SVFs from IWAT were efficiently knocked down after a 3-day infection of mouse Irx3 lentiviral shRNA ( n = 3 for each group). (C–D) (C) Oil Red O staining and (D) TG concentration of the mature beige adipocytes under eight-day differentiation ( n = 6 for each group). (E) Relative mRNA expression of Ucp1 , Cidea , Pgc-1α , C/ebpβ , and Prdm16 at different time points during beige adipocyte differentiation from IWAT SVFs ( n = 3–4 for different groups). (F–G) (F) Relative mRNA ( n = 4) and (G) protein expression of brown adipocyte marker genes of the induced beige adipocytes from mouse IWAT SVTs for eight days with the infection of mouse Irx3 lentiviral shRNA, with or without CL-316,243 activation. (H) Relative mRNA expression of adipocyte differentiation-related genes in beige adipocytes ( n = 4). (I) Relative mRNA expression of the indicated genes in the induced brown adipocytes from mouse BAT SVFs for eight days, with the infection of mouse Irx3 lentiviral shRNA ( n = 4). (J) Relative mRNA levels of IRX3 in SVFs from human sWAT after a 3-day infection of human IRX3 lentiviral shRNA (left), and the mRNA levels of brown adipocyte marker genes in the SVFs under 2-day differentiation ( n = 4). (K) OCR measurement of the beige adipocytes under 5-day differentiation, with the infection of mouse Irx3 lentiviral shRNA ( n = 5). (L–N) (L) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector or IRX3 for 48 h. (M) Schematic diagram of the mutant mouse Ucp1 promoter deleting ACATGTGT among the − 3470 to − 3463 bp region proximal to the TSS of the Ucp1 gene. (N) Transcriptional activity of wild-type or mutant Ucp1 promoter was measured with IRX3 overexpression. (O) qPCR of Ucp1 promoter binding region which was recruited by Irx3 antibody in 8-day induced beige adipocytes, as analyzed by ChIP. Fold enrichment of Ucp1 promoter was given ( n = 3). For the measurement of luciferase activity, cells were seeded on 24-well plate and transfected with 800 ng pEGFP-C1 vector or IRX3, 200 ng Ucp1 promoter construct, and 1 ng pRL-SV40, followed by the harvest for luciferase activity assessment using a dual-luciferase reporter assay system (Promega). Luciferase activity was corrected for Renilla luciferase activity ( n = 3–4 for each group). For qPCR data, mRNA expression was normalized to 36b4 for mouse genes and βACTIN for human genes. SVF cells were isolated from IWAT and BAT of C57BL/6J mice or human sWAT wherever mentioned, and IRX3 shRNA were introduced to cells 24 h after seeding. Data were presented as mean ± s.e.m. * P

    Journal: EBioMedicine

    Article Title: IRX3 Promotes the Browning of White Adipocytes and Its Rare Variants are Associated with Human Obesity Risk

    doi: 10.1016/j.ebiom.2017.09.010

    Figure Lengend Snippet: Knockdown of Irx3 repressed the differentiation of SVFs toward beige adipocytes in vitro. (A–B) (A) Relative mRNA and (B) protein levels of Irx3 in SVFs from IWAT were efficiently knocked down after a 3-day infection of mouse Irx3 lentiviral shRNA ( n = 3 for each group). (C–D) (C) Oil Red O staining and (D) TG concentration of the mature beige adipocytes under eight-day differentiation ( n = 6 for each group). (E) Relative mRNA expression of Ucp1 , Cidea , Pgc-1α , C/ebpβ , and Prdm16 at different time points during beige adipocyte differentiation from IWAT SVFs ( n = 3–4 for different groups). (F–G) (F) Relative mRNA ( n = 4) and (G) protein expression of brown adipocyte marker genes of the induced beige adipocytes from mouse IWAT SVTs for eight days with the infection of mouse Irx3 lentiviral shRNA, with or without CL-316,243 activation. (H) Relative mRNA expression of adipocyte differentiation-related genes in beige adipocytes ( n = 4). (I) Relative mRNA expression of the indicated genes in the induced brown adipocytes from mouse BAT SVFs for eight days, with the infection of mouse Irx3 lentiviral shRNA ( n = 4). (J) Relative mRNA levels of IRX3 in SVFs from human sWAT after a 3-day infection of human IRX3 lentiviral shRNA (left), and the mRNA levels of brown adipocyte marker genes in the SVFs under 2-day differentiation ( n = 4). (K) OCR measurement of the beige adipocytes under 5-day differentiation, with the infection of mouse Irx3 lentiviral shRNA ( n = 5). (L–N) (L) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector or IRX3 for 48 h. (M) Schematic diagram of the mutant mouse Ucp1 promoter deleting ACATGTGT among the − 3470 to − 3463 bp region proximal to the TSS of the Ucp1 gene. (N) Transcriptional activity of wild-type or mutant Ucp1 promoter was measured with IRX3 overexpression. (O) qPCR of Ucp1 promoter binding region which was recruited by Irx3 antibody in 8-day induced beige adipocytes, as analyzed by ChIP. Fold enrichment of Ucp1 promoter was given ( n = 3). For the measurement of luciferase activity, cells were seeded on 24-well plate and transfected with 800 ng pEGFP-C1 vector or IRX3, 200 ng Ucp1 promoter construct, and 1 ng pRL-SV40, followed by the harvest for luciferase activity assessment using a dual-luciferase reporter assay system (Promega). Luciferase activity was corrected for Renilla luciferase activity ( n = 3–4 for each group). For qPCR data, mRNA expression was normalized to 36b4 for mouse genes and βACTIN for human genes. SVF cells were isolated from IWAT and BAT of C57BL/6J mice or human sWAT wherever mentioned, and IRX3 shRNA were introduced to cells 24 h after seeding. Data were presented as mean ± s.e.m. * P

    Article Snippet: 2.14 Chromatin Immunoprecipitation Assay Chromatin immunoprecipitation (ChIP) assay was carried out to investigate the interaction between IRX3 and the Ucp1 promoter using a commercial kit (Millipore, 17–371) in accordance with the manufacturer's instructions.

    Techniques: In Vitro, Infection, shRNA, Staining, Concentration Assay, Expressing, Pyrolysis Gas Chromatography, Marker, Activation Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Over Expression, Real-time Polymerase Chain Reaction, Binding Assay, Chromatin Immunoprecipitation, Construct, Reporter Assay, Isolation, Mouse Assay

    The expression of Irx3 was correlated with the beige/brown adipocyte associated genes in vitro. (A–G) In fully differentiated beige/brown adipocytes from preadipocytes isolated from IWAT, EWAT, and BAT of C57BL/6J and 129/Sv mice, Ucp1 mRNA expression was correlated with Pgc-1α (A) and Dio2 (B). Irx3 mRNA expression showed positive correlation with mRNA levels of (C) Ucp1 , (D) Pgc-1α , (E) Dio2 , and (F) Cox7a1 . Irx3 mRNA expression showed no correlation with Leptin (G). Cells from C57BL/6J were shown in red, and from 129/Sv were in blue. Cells from IWAT, EWAT, and BAT were shown as dot, triangle, and square, respectively. The correlation was analyzed using ΔCT. (H-J) (H) Irx3 (I) Ucp1 and (J) Pparγ mRNA expression during the time course of beige adipocyte differentiation cultures ( n = 3). Quantitative amounts of gene expression were normalized to the housekeeping gene 36b4 . (K–L) Irx3 and Ucp1 protein levels at different time points during beige adipocyte differentiation from IWAT SVFs (K), during brown adipocyte differentiation from BAT SVFs (L). (M) Protein expression of Irx3 and Ap2 at different time points during white adipocyte differentiation from IWAT SVFs. (N) Intracellular location of Ucp1 and Irx3 protein in the fully differentiated beige adipocytes. Ucp1 protein, Irx3 protein and nucleus were indicated in green, red, and blue, respectively. Scar bar, 20 μm. Data were presented as mean ± s.e.m. * P

    Journal: EBioMedicine

    Article Title: IRX3 Promotes the Browning of White Adipocytes and Its Rare Variants are Associated with Human Obesity Risk

    doi: 10.1016/j.ebiom.2017.09.010

    Figure Lengend Snippet: The expression of Irx3 was correlated with the beige/brown adipocyte associated genes in vitro. (A–G) In fully differentiated beige/brown adipocytes from preadipocytes isolated from IWAT, EWAT, and BAT of C57BL/6J and 129/Sv mice, Ucp1 mRNA expression was correlated with Pgc-1α (A) and Dio2 (B). Irx3 mRNA expression showed positive correlation with mRNA levels of (C) Ucp1 , (D) Pgc-1α , (E) Dio2 , and (F) Cox7a1 . Irx3 mRNA expression showed no correlation with Leptin (G). Cells from C57BL/6J were shown in red, and from 129/Sv were in blue. Cells from IWAT, EWAT, and BAT were shown as dot, triangle, and square, respectively. The correlation was analyzed using ΔCT. (H-J) (H) Irx3 (I) Ucp1 and (J) Pparγ mRNA expression during the time course of beige adipocyte differentiation cultures ( n = 3). Quantitative amounts of gene expression were normalized to the housekeeping gene 36b4 . (K–L) Irx3 and Ucp1 protein levels at different time points during beige adipocyte differentiation from IWAT SVFs (K), during brown adipocyte differentiation from BAT SVFs (L). (M) Protein expression of Irx3 and Ap2 at different time points during white adipocyte differentiation from IWAT SVFs. (N) Intracellular location of Ucp1 and Irx3 protein in the fully differentiated beige adipocytes. Ucp1 protein, Irx3 protein and nucleus were indicated in green, red, and blue, respectively. Scar bar, 20 μm. Data were presented as mean ± s.e.m. * P

    Article Snippet: 2.14 Chromatin Immunoprecipitation Assay Chromatin immunoprecipitation (ChIP) assay was carried out to investigate the interaction between IRX3 and the Ucp1 promoter using a commercial kit (Millipore, 17–371) in accordance with the manufacturer's instructions.

    Techniques: Expressing, In Vitro, Isolation, Mouse Assay, Pyrolysis Gas Chromatography

    The association of IRX3 and human obesity. (A) Relative mRNA levels of IRX3 in oWAT or sWAT from normal weight ( n = 9) and obese subjects ( n = 21) measured by qPCR. Gene expression was normalized to βACTIN . (B) Comparison of the frequency of the IRX3 rare variants in normal weight subjects and obese patients. (C) Sequence conservation analysis of the IRX3 orthologs related to variants identified in obese subjects and controls. Mutant sites are shown with a red box. Dark gray represents residues that are completely conserved, mild gray partially conserved, and light gray shows similar residues. (D) Protein expression levels of WT and mutant IRX3 in HEK293T cells. GFP antibody was used to indicate IRX3, and actin antibody served as a loading control. (E) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector, WT or mutant IRX3 for 48 h ( n = 3). Normalized luciferase activities are shown as fold change and compared to WT IRX3 ( n = 4). oWAT, omental white adipose tissue. sWAT, subcutaneous white adipose tissue. Data were presented as mean ± s.e.m. * P

    Journal: EBioMedicine

    Article Title: IRX3 Promotes the Browning of White Adipocytes and Its Rare Variants are Associated with Human Obesity Risk

    doi: 10.1016/j.ebiom.2017.09.010

    Figure Lengend Snippet: The association of IRX3 and human obesity. (A) Relative mRNA levels of IRX3 in oWAT or sWAT from normal weight ( n = 9) and obese subjects ( n = 21) measured by qPCR. Gene expression was normalized to βACTIN . (B) Comparison of the frequency of the IRX3 rare variants in normal weight subjects and obese patients. (C) Sequence conservation analysis of the IRX3 orthologs related to variants identified in obese subjects and controls. Mutant sites are shown with a red box. Dark gray represents residues that are completely conserved, mild gray partially conserved, and light gray shows similar residues. (D) Protein expression levels of WT and mutant IRX3 in HEK293T cells. GFP antibody was used to indicate IRX3, and actin antibody served as a loading control. (E) Ucp1 promoter-luciferase reporter activity was measured in HEK293T cells transfected with pEGFP-C1 vector, WT or mutant IRX3 for 48 h ( n = 3). Normalized luciferase activities are shown as fold change and compared to WT IRX3 ( n = 4). oWAT, omental white adipose tissue. sWAT, subcutaneous white adipose tissue. Data were presented as mean ± s.e.m. * P

    Article Snippet: 2.14 Chromatin Immunoprecipitation Assay Chromatin immunoprecipitation (ChIP) assay was carried out to investigate the interaction between IRX3 and the Ucp1 promoter using a commercial kit (Millipore, 17–371) in accordance with the manufacturer's instructions.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Sequencing, Mutagenesis, Luciferase, Activity Assay, Transfection, Plasmid Preparation

    IRX3/Irx3 expression is induced in browning adipose tissues. (A) Relative mRNA levels of Ucp1 and Irx3 in IWAT from mice under cold stress (4 °C) for one week ( n = 6) and at 25 °C ( n = 8). (B–C) Relative mRNA levels of Ucp1 and Irx3 in IWAT (B) and BAT (C) from mice subjected to PBS or CL-316,243 for 10 days. For A–C, gene expression was normalized to 36b4 . (D–E) Protein levels of Ucp1 and Irx3 (left) and the quantification value of Irx3 relative to Hsp90 (right) in IWAT (D) and BAT (E) in the mice subjected to PBS or CL-316,243 treatment. (F) Representative images of immunohistochemical staining in IWAT from mice subjected to PBS or CL-316,243 treatment (scale bar, 50 μm for the upper and middle panels, and 100 μm for the bottom panel). (G) PET-CT, gross image, HE staining (scale bar, 20 μm), and electron microscopy (scale bar, 5 μm) of the browning WAT from pheochromocytoma patients. (H) Representative images of immunohistochemical staining in oWAT from pheochromocytoma patients (scale bar, 100 μm for the upper panel, and 50 μm for the bottom panel). Data were presented as mean ± s.e.m. * P

    Journal: EBioMedicine

    Article Title: IRX3 Promotes the Browning of White Adipocytes and Its Rare Variants are Associated with Human Obesity Risk

    doi: 10.1016/j.ebiom.2017.09.010

    Figure Lengend Snippet: IRX3/Irx3 expression is induced in browning adipose tissues. (A) Relative mRNA levels of Ucp1 and Irx3 in IWAT from mice under cold stress (4 °C) for one week ( n = 6) and at 25 °C ( n = 8). (B–C) Relative mRNA levels of Ucp1 and Irx3 in IWAT (B) and BAT (C) from mice subjected to PBS or CL-316,243 for 10 days. For A–C, gene expression was normalized to 36b4 . (D–E) Protein levels of Ucp1 and Irx3 (left) and the quantification value of Irx3 relative to Hsp90 (right) in IWAT (D) and BAT (E) in the mice subjected to PBS or CL-316,243 treatment. (F) Representative images of immunohistochemical staining in IWAT from mice subjected to PBS or CL-316,243 treatment (scale bar, 50 μm for the upper and middle panels, and 100 μm for the bottom panel). (G) PET-CT, gross image, HE staining (scale bar, 20 μm), and electron microscopy (scale bar, 5 μm) of the browning WAT from pheochromocytoma patients. (H) Representative images of immunohistochemical staining in oWAT from pheochromocytoma patients (scale bar, 100 μm for the upper panel, and 50 μm for the bottom panel). Data were presented as mean ± s.e.m. * P

    Article Snippet: 2.14 Chromatin Immunoprecipitation Assay Chromatin immunoprecipitation (ChIP) assay was carried out to investigate the interaction between IRX3 and the Ucp1 promoter using a commercial kit (Millipore, 17–371) in accordance with the manufacturer's instructions.

    Techniques: Expressing, Mouse Assay, Immunohistochemistry, Staining, Positron Emission Tomography, Electron Microscopy

    Quercetin promoted translocation of PGC‐1α from cytoplasm to nucleus and enhanced PGC‐1α binding. ( A‐D ) The representative photomicrographs showing PGC‐1α immunohistochemistry of tissue from different group 24 hrs after TBI. ( E, H ) The nuclear PGC‐1α expression after quercetin treatment in mice with TBI, as measured by Western blot. ( F, I ) The cytoplasmic protein PGC‐1α expression after quercetin treatment in mice with TBI, as measured by Western blot. ( G, J ) The total protein PGC‐1α expression after melatonin treatment in mice with TBI, as measured by Western blot. Bars represent the mean ± S.E.M. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Neuroprotection by quercetin via mitochondrial function adaptation in traumatic brain injury: PGC‐1α pathway as a potential mechanism

    doi: 10.1111/jcmm.13313

    Figure Lengend Snippet: Quercetin promoted translocation of PGC‐1α from cytoplasm to nucleus and enhanced PGC‐1α binding. ( A‐D ) The representative photomicrographs showing PGC‐1α immunohistochemistry of tissue from different group 24 hrs after TBI. ( E, H ) The nuclear PGC‐1α expression after quercetin treatment in mice with TBI, as measured by Western blot. ( F, I ) The cytoplasmic protein PGC‐1α expression after quercetin treatment in mice with TBI, as measured by Western blot. ( G, J ) The total protein PGC‐1α expression after melatonin treatment in mice with TBI, as measured by Western blot. Bars represent the mean ± S.E.M. * P

    Article Snippet: The nuclear protein was incubated overnight at 4°C with PGC‐1α (1:1000 diluted, rabbit; Millipore, Billerica, MA, USA) and histone H3 (1:1000 diluted, rabbit; Cell Signaling Technology, Beverly, MA, USA) in blocking buffer.

    Techniques: Translocation Assay, Pyrolysis Gas Chromatography, Binding Assay, Immunohistochemistry, Expressing, Mouse Assay, Western Blot

    MT prevents tBHP inhibition of Akt function in vitro. Adult cardiomyocytes from WT and MT-TG mice were directly exposed to tBHP (100 μmol/L) for 2 h or LPS (10 µg/mL) for 4 h and then stimulated with or without insulin (INS; 100 nmol/L) for 15 min. Cells were collected, and the phosphorylation of total Akt ( A and B ), Akt2 ( A , C , I , and J ), GSK-3β ( A , D , I , and K ), and GS ( A , E , I , and L ) and expression of HK II ( A , F , I , and M ), PGC-1α ( A , G , I , and N ), and TRB3 ( A , H , I , and O ) was detected by Western blot. Results were normalized to WT control. Three independent experiments were performed. Data shown in graphs represent the means ± SD. * P

    Journal: Diabetes

    Article Title: Metallothionein Preserves Akt2 Activity and Cardiac Function via Inhibiting TRB3 in Diabetic Hearts

    doi: 10.2337/db17-0219

    Figure Lengend Snippet: MT prevents tBHP inhibition of Akt function in vitro. Adult cardiomyocytes from WT and MT-TG mice were directly exposed to tBHP (100 μmol/L) for 2 h or LPS (10 µg/mL) for 4 h and then stimulated with or without insulin (INS; 100 nmol/L) for 15 min. Cells were collected, and the phosphorylation of total Akt ( A and B ), Akt2 ( A , C , I , and J ), GSK-3β ( A , D , I , and K ), and GS ( A , E , I , and L ) and expression of HK II ( A , F , I , and M ), PGC-1α ( A , G , I , and N ), and TRB3 ( A , H , I , and O ) was detected by Western blot. Results were normalized to WT control. Three independent experiments were performed. Data shown in graphs represent the means ± SD. * P

    Article Snippet: TRB3 is an Akt-negative regulator that directly binds to and blocks Akt phosphorylation ( , , ).

    Techniques: Inhibition, In Vitro, Mouse Assay, Expressing, Pyrolysis Gas Chromatography, Western Blot

    MT prevents diabetic downregulation of cardiac Akt functions along with inhibition of TRB3 in STZ-induced type 1 diabetes. At 3 months after diabetes onset, WT and MT-TG mice were sacrificed, and cardiac tissue was used to analyze the phosphorylation of total Akt ( A and B ), Akt1 ( A and C ), Akt2 ( A and D ), GSK-3β ( A and E ), and GS ( A and F ) and the expression of HK II ( A and G ), PGC-1α ( A and H ), and TRB3 ( A and I ) by Western blot. Results were normalized to the WT controls. GAPDH was used as loading control. n = 7 for each group. Data shown in graphs represent the means ± SD. * P

    Journal: Diabetes

    Article Title: Metallothionein Preserves Akt2 Activity and Cardiac Function via Inhibiting TRB3 in Diabetic Hearts

    doi: 10.2337/db17-0219

    Figure Lengend Snippet: MT prevents diabetic downregulation of cardiac Akt functions along with inhibition of TRB3 in STZ-induced type 1 diabetes. At 3 months after diabetes onset, WT and MT-TG mice were sacrificed, and cardiac tissue was used to analyze the phosphorylation of total Akt ( A and B ), Akt1 ( A and C ), Akt2 ( A and D ), GSK-3β ( A and E ), and GS ( A and F ) and the expression of HK II ( A and G ), PGC-1α ( A and H ), and TRB3 ( A and I ) by Western blot. Results were normalized to the WT controls. GAPDH was used as loading control. n = 7 for each group. Data shown in graphs represent the means ± SD. * P

    Article Snippet: TRB3 is an Akt-negative regulator that directly binds to and blocks Akt phosphorylation ( , , ).

    Techniques: Inhibition, Mouse Assay, Expressing, Pyrolysis Gas Chromatography, Western Blot