ps3-000 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 98
    Thermo Fisher anti atp synthase β subunit
    Anti Atp Synthase β Subunit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti atp synthase β subunit/product/Thermo Fisher
    Average 98 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    anti atp synthase β subunit - by Bioz Stars, 2020-09
    98/100 stars
      Buy from Supplier

    96
    Millipore anti ogt antibody
    Loss of skeletal muscle <t>OGT</t> alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) <t>UDP-GlcNAc</t> levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Anti Ogt Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti ogt antibody/product/Millipore
    Average 96 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    anti ogt antibody - by Bioz Stars, 2020-09
    96/100 stars
      Buy from Supplier

    91
    Cell Signaling Technology Inc akt 1 2 3
    Loss of skeletal muscle <t>OGT</t> alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) <t>UDP-GlcNAc</t> levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Akt 1 2 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/akt 1 2 3/product/Cell Signaling Technology Inc
    Average 91 stars, based on 32 article reviews
    Price from $9.99 to $1999.99
    akt 1 2 3 - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    90
    Cell Signaling Technology Inc akt pt308
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of <t>Akt</t> and Akt phosphorylation of pS473 and <t>pT308</t> in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Akt Pt308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/akt pt308/product/Cell Signaling Technology Inc
    Average 90 stars, based on 77 article reviews
    Price from $9.99 to $1999.99
    akt pt308 - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    92
    Cell Signaling Technology Inc akt ps473
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of <t>Akt</t> and Akt phosphorylation of <t>pS473</t> and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Akt Ps473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 289 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/akt ps473/product/Cell Signaling Technology Inc
    Average 92 stars, based on 289 article reviews
    Price from $9.99 to $1999.99
    akt ps473 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    94
    Santa Cruz Biotechnology akt1
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of <t>Akt1</t> in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Akt1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 991 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/akt1/product/Santa Cruz Biotechnology
    Average 94 stars, based on 991 article reviews
    Price from $9.99 to $1999.99
    akt1 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    94
    Abcam anti succinate dehydrogenase a subunit sdha
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of <t>Akt1</t> in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Anti Succinate Dehydrogenase A Subunit Sdha, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti succinate dehydrogenase a subunit sdha/product/Abcam
    Average 94 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    anti succinate dehydrogenase a subunit sdha - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    90
    Abcam anti o glcnac
    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and <t>anti-O-GlcNAc</t> antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p
    Anti O Glcnac, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 139 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti o glcnac/product/Abcam
    Average 90 stars, based on 139 article reviews
    Price from $9.99 to $1999.99
    anti o glcnac - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    96
    Cell Signaling Technology Inc insulin receptor β
    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and <t>anti-O-GlcNAc</t> antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p
    Insulin Receptor β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/insulin receptor β/product/Cell Signaling Technology Inc
    Average 96 stars, based on 85 article reviews
    Price from $9.99 to $1999.99
    insulin receptor β - by Bioz Stars, 2020-09
    96/100 stars
      Buy from Supplier

    93
    Thermo Fisher anti glucose transporter 4
    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and <t>anti-O-GlcNAc</t> antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p
    Anti Glucose Transporter 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti glucose transporter 4/product/Thermo Fisher
    Average 93 stars, based on 52 article reviews
    Price from $9.99 to $1999.99
    anti glucose transporter 4 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc hexokinase ii
    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and <t>anti-O-GlcNAc</t> antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p
    Hexokinase Ii, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 197 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hexokinase ii/product/Cell Signaling Technology Inc
    Average 94 stars, based on 197 article reviews
    Price from $9.99 to $1999.99
    hexokinase ii - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc atgl
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of <t>HSL</t> and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of <t>ATGL</t> in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Atgl, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 653 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/atgl/product/Cell Signaling Technology Inc
    Average 99 stars, based on 653 article reviews
    Price from $9.99 to $1999.99
    atgl - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    gfat1  (Abcam)
    92
    Abcam gfat1
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of <t>GFAT1</t> in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Gfat1, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfat1/product/Abcam
    Average 92 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    gfat1 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    90
    Cell Signaling Technology Inc hsl ps563
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of <t>HSL</t> and HSL <t>pS563</t> in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Hsl Ps563, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hsl ps563/product/Cell Signaling Technology Inc
    Average 90 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    hsl ps563 - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc hsl
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of <t>HSL</t> and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of <t>ATGL</t> in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Hsl, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 667 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hsl/product/Cell Signaling Technology Inc
    Average 99 stars, based on 667 article reviews
    Price from $9.99 to $1999.99
    hsl - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc irs1
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of <t>HSL</t> and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of <t>ATGL</t> in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Irs1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 579 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/irs1/product/Cell Signaling Technology Inc
    Average 99 stars, based on 579 article reviews
    Price from $9.99 to $1999.99
    irs1 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    Tektronix keithley 2400 source meter
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of <t>HSL</t> and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of <t>ATGL</t> in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Keithley 2400 Source Meter, supplied by Tektronix, used in various techniques. Bioz Stars score: 91/100, based on 221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/keithley 2400 source meter/product/Tektronix
    Average 91 stars, based on 221 article reviews
    Price from $9.99 to $1999.99
    keithley 2400 source meter - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    93
    Cell Signaling Technology Inc o glcnac
    Loss of skeletal muscle <t>OGT</t> alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) <t>UDP-GlcNAc</t> levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    O Glcnac, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/o glcnac/product/Cell Signaling Technology Inc
    Average 93 stars, based on 59 article reviews
    Price from $9.99 to $1999.99
    o glcnac - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    oga  (Abcam)
    90
    Abcam oga
    Loss of skeletal muscle <t>OGT</t> alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) <t>UDP-GlcNAc</t> levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Oga, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oga/product/Abcam
    Average 90 stars, based on 81 article reviews
    Price from $9.99 to $1999.99
    oga - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc ogt
    Loss of skeletal muscle <t>OGT</t> alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) <t>UDP-GlcNAc</t> levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Ogt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ogt/product/Cell Signaling Technology Inc
    Average 94 stars, based on 67 article reviews
    Price from $9.99 to $1999.99
    ogt - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    90
    Cell Signaling Technology Inc pdh
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase <t>(PDH)</t> activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at <t>pS293</t> and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Pdh, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 214 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pdh/product/Cell Signaling Technology Inc
    Average 90 stars, based on 214 article reviews
    Price from $9.99 to $1999.99
    pdh - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    94
    Santa Cruz Biotechnology pfk1
    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase <t>(PDH)</t> activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at <t>pS293</t> and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p
    Pfk1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pfk1/product/Santa Cruz Biotechnology
    Average 94 stars, based on 65 article reviews
    Price from $9.99 to $1999.99
    pfk1 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    Millipore tbc1d4
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of <t>TBC1D4</t> and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Tbc1d4, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tbc1d4/product/Millipore
    Average 99 stars, based on 47 article reviews
    Price from $9.99 to $1999.99
    tbc1d4 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Millipore acc ps79 257
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of <t>TBC1D4</t> and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Acc Ps79 257, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/acc ps79 257/product/Millipore
    Average 92 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    acc ps79 257 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Thermo Fisher mem
    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of <t>TBC1D4</t> and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p
    Mem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 9423 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mem/product/Thermo Fisher
    Average 92 stars, based on 9423 article reviews
    Price from $9.99 to $1999.99
    mem - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    Image Search Results


    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Inducible knockout of OGT in skeletal muscle recapitulates the mKO mouse. OGT knockout was induced when mice were 5 months old by feeding doxycycline food for two weeks. Subsequently, mice were fed normal chow for 6 months before analysis. (A) Muscle from WT and imKO mice were immunoblotted with anti-OGT and -O-GlcNAc antibodies respectively. Equal amount of total protein was loaded across all the lanes. (B – F) Body weight (B) , muscle mass (C) , fat mass (D) , fat depots (E) , and energy expenditure (EE) (F) were assessed. (G – H) Glucose tolerance test (GTT) (G) and insulin tolerance test (ITT) (H) . Data represent means ± SEM from n = 10 male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Inducible knockout of OGT in skeletal muscle recapitulates the mKO mouse. OGT knockout was induced when mice were 5 months old by feeding doxycycline food for two weeks. Subsequently, mice were fed normal chow for 6 months before analysis. (A) Muscle from WT and imKO mice were immunoblotted with anti-OGT and -O-GlcNAc antibodies respectively. Equal amount of total protein was loaded across all the lanes. (B – F) Body weight (B) , muscle mass (C) , fat mass (D) , fat depots (E) , and energy expenditure (EE) (F) were assessed. (G – H) Glucose tolerance test (GTT) (G) and insulin tolerance test (ITT) (H) . Data represent means ± SEM from n = 10 male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Mouse Assay

    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Expressing, Mouse Assay, Staining, Cell Culture, Isolation, Immunoprecipitation, Chromatin Immunoprecipitation, Quantitative RT-PCR, Sequencing

    Mice lacking OGT in skeletal muscle exhibit reduced fat mass but normal skeletal muscle morphology and contractility. (A) O-GlcNAc levels in human skeletal muscles of lean, obese, and type 2 diabetic individuals at basal (B) and after a hyperinsulinemic euglycemic clamp (Insulin (I)). Lane intensities were quantified between 15 and 300 kDa. Sample size of 8–10 people per group. Data are mean ± SEM and analyzed by two-way repeated measures ANOVA; ** indicates p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Mice lacking OGT in skeletal muscle exhibit reduced fat mass but normal skeletal muscle morphology and contractility. (A) O-GlcNAc levels in human skeletal muscles of lean, obese, and type 2 diabetic individuals at basal (B) and after a hyperinsulinemic euglycemic clamp (Insulin (I)). Lane intensities were quantified between 15 and 300 kDa. Sample size of 8–10 people per group. Data are mean ± SEM and analyzed by two-way repeated measures ANOVA; ** indicates p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay

    Loss of skeletal muscle OGT increases type I myosin heavy chain-containing fibers, but impairs maximal oxidative capacity. (A) Relative mRNA expression of myosin heavy chain (MyHC) I ( Myh7 ), MyHC IIa ( Myh2 ), MyHC IIx ( Myh1 ), and MyHC IIb ( Myh4 ) in the gastrocnemius muscle. Inset, immunoreactivity of MyHC I in the gastrocnemius muscles of WT and mKO muscle. (B) Mitochondrial abundance. Left: three-dimensional confocal images showing the difference in mitochondria in gastrocnemius muscle cross-sections of WT and mKO stained with succinate dehydrogenase antibody (left panel). Bar, 5 μm. Right: mitochondrial DNA (mtDNA) abundance in the gastrocnemius muscles of WT and mKO mice. Mt–Co1 DNA was measured using qPCR and normalized to genomic Nrip1 DNA. (C – D) Citrate synthase (CS, C) and malate dehydrogenase (MDH, D) activities in isolated mitochondria (Mito), and red (R.) and white (W.) portions of the gastrocnemius muscles of WT and mKO mice. (E) Abundance of mitochondrial complexes in gastrocnemius from clamped mice. (F) Mitochondria were isolated from WT and mKO gastrocnemius muscle, and oxygen consumption was measured using Seahorse. Respiration control ratio of state 3/state 4 was calculated based on oxygen consumption rate (OCR) in each state. Data represent means ± SEM from n = 10, 16-week-old male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT increases type I myosin heavy chain-containing fibers, but impairs maximal oxidative capacity. (A) Relative mRNA expression of myosin heavy chain (MyHC) I ( Myh7 ), MyHC IIa ( Myh2 ), MyHC IIx ( Myh1 ), and MyHC IIb ( Myh4 ) in the gastrocnemius muscle. Inset, immunoreactivity of MyHC I in the gastrocnemius muscles of WT and mKO muscle. (B) Mitochondrial abundance. Left: three-dimensional confocal images showing the difference in mitochondria in gastrocnemius muscle cross-sections of WT and mKO stained with succinate dehydrogenase antibody (left panel). Bar, 5 μm. Right: mitochondrial DNA (mtDNA) abundance in the gastrocnemius muscles of WT and mKO mice. Mt–Co1 DNA was measured using qPCR and normalized to genomic Nrip1 DNA. (C – D) Citrate synthase (CS, C) and malate dehydrogenase (MDH, D) activities in isolated mitochondria (Mito), and red (R.) and white (W.) portions of the gastrocnemius muscles of WT and mKO mice. (E) Abundance of mitochondrial complexes in gastrocnemius from clamped mice. (F) Mitochondria were isolated from WT and mKO gastrocnemius muscle, and oxygen consumption was measured using Seahorse. Respiration control ratio of state 3/state 4 was calculated based on oxygen consumption rate (OCR) in each state. Data represent means ± SEM from n = 10, 16-week-old male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Expressing, Staining, Mouse Assay, Real-time Polymerase Chain Reaction, Isolation

    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay

    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay

    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay

    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Expressing, Mouse Assay, Staining, Cell Culture, Isolation, Immunoprecipitation, Chromatin Immunoprecipitation, Quantitative RT-PCR, Sequencing

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Inducible knockout of OGT in skeletal muscle recapitulates the mKO mouse. OGT knockout was induced when mice were 5 months old by feeding doxycycline food for two weeks. Subsequently, mice were fed normal chow for 6 months before analysis. (A) Muscle from WT and imKO mice were immunoblotted with anti-OGT and -O-GlcNAc antibodies respectively. Equal amount of total protein was loaded across all the lanes. (B – F) Body weight (B) , muscle mass (C) , fat mass (D) , fat depots (E) , and energy expenditure (EE) (F) were assessed. (G – H) Glucose tolerance test (GTT) (G) and insulin tolerance test (ITT) (H) . Data represent means ± SEM from n = 10 male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Inducible knockout of OGT in skeletal muscle recapitulates the mKO mouse. OGT knockout was induced when mice were 5 months old by feeding doxycycline food for two weeks. Subsequently, mice were fed normal chow for 6 months before analysis. (A) Muscle from WT and imKO mice were immunoblotted with anti-OGT and -O-GlcNAc antibodies respectively. Equal amount of total protein was loaded across all the lanes. (B – F) Body weight (B) , muscle mass (C) , fat mass (D) , fat depots (E) , and energy expenditure (EE) (F) were assessed. (G – H) Glucose tolerance test (GTT) (G) and insulin tolerance test (ITT) (H) . Data represent means ± SEM from n = 10 male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Mouse Assay

    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Expressing, Mouse Assay, Staining, Cell Culture, Isolation, Immunoprecipitation, Chromatin Immunoprecipitation, Quantitative RT-PCR, Sequencing

    Mice lacking OGT in skeletal muscle exhibit reduced fat mass but normal skeletal muscle morphology and contractility. (A) O-GlcNAc levels in human skeletal muscles of lean, obese, and type 2 diabetic individuals at basal (B) and after a hyperinsulinemic euglycemic clamp (Insulin (I)). Lane intensities were quantified between 15 and 300 kDa. Sample size of 8–10 people per group. Data are mean ± SEM and analyzed by two-way repeated measures ANOVA; ** indicates p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Mice lacking OGT in skeletal muscle exhibit reduced fat mass but normal skeletal muscle morphology and contractility. (A) O-GlcNAc levels in human skeletal muscles of lean, obese, and type 2 diabetic individuals at basal (B) and after a hyperinsulinemic euglycemic clamp (Insulin (I)). Lane intensities were quantified between 15 and 300 kDa. Sample size of 8–10 people per group. Data are mean ± SEM and analyzed by two-way repeated measures ANOVA; ** indicates p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Inducible knockout of OGT in skeletal muscle recapitulates the mKO mouse. OGT knockout was induced when mice were 5 months old by feeding doxycycline food for two weeks. Subsequently, mice were fed normal chow for 6 months before analysis. (A) Muscle from WT and imKO mice were immunoblotted with anti-OGT and -O-GlcNAc antibodies respectively. Equal amount of total protein was loaded across all the lanes. (B – F) Body weight (B) , muscle mass (C) , fat mass (D) , fat depots (E) , and energy expenditure (EE) (F) were assessed. (G – H) Glucose tolerance test (GTT) (G) and insulin tolerance test (ITT) (H) . Data represent means ± SEM from n = 10 male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Inducible knockout of OGT in skeletal muscle recapitulates the mKO mouse. OGT knockout was induced when mice were 5 months old by feeding doxycycline food for two weeks. Subsequently, mice were fed normal chow for 6 months before analysis. (A) Muscle from WT and imKO mice were immunoblotted with anti-OGT and -O-GlcNAc antibodies respectively. Equal amount of total protein was loaded across all the lanes. (B – F) Body weight (B) , muscle mass (C) , fat mass (D) , fat depots (E) , and energy expenditure (EE) (F) were assessed. (G – H) Glucose tolerance test (GTT) (G) and insulin tolerance test (ITT) (H) . Data represent means ± SEM from n = 10 male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Mouse Assay

    OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: OGT knockout results in augmented Interleukin-15 production in skeletal muscle. (A) Expression of myokines in the gastrocnemius muscles of WT and mKO mice fed HFD for 22 wks (B) IL-15 mRNA expression in skeletal muscles from clamped mice (n = 8–10, 18-week-old male mice). (C) Serum IL-15 levels in WT and mKO mice fed HFD for 22 wks (n = 10). (D) Oil red-O staining of cultured stromal vascular fraction cells isolated from mouse IngW and induced to differentiate in the presence or absence of gastrocnemius muscle extracts from WT (top row) and mKO (bottom row) mice with (right) or without (left) IL15 neutralizing antibodies. Scale bar, 1 mm. (E) Expression of lipogenic genes in IngW (n = 10). (F) Immunoprecipitation was performed using anti-EZH2 and anti-O-GlcNAc antibodies respectively, and immunoblotted with O-GlcNAc and EZH2 antibodies respectively. (G – I) Chromatin immunoprecipitation using antibodies against O-GlcNAc (G), EZH2 (H), and H3K27me3 (I), respectively. Quantitative RT-PCR was used to quantify IL-15 promoter sequence as detailed in Methods. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Knock-Out, Expressing, Mouse Assay, Staining, Cell Culture, Isolation, Immunoprecipitation, Chromatin Immunoprecipitation, Quantitative RT-PCR, Sequencing

    Mice lacking OGT in skeletal muscle exhibit reduced fat mass but normal skeletal muscle morphology and contractility. (A) O-GlcNAc levels in human skeletal muscles of lean, obese, and type 2 diabetic individuals at basal (B) and after a hyperinsulinemic euglycemic clamp (Insulin (I)). Lane intensities were quantified between 15 and 300 kDa. Sample size of 8–10 people per group. Data are mean ± SEM and analyzed by two-way repeated measures ANOVA; ** indicates p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Mice lacking OGT in skeletal muscle exhibit reduced fat mass but normal skeletal muscle morphology and contractility. (A) O-GlcNAc levels in human skeletal muscles of lean, obese, and type 2 diabetic individuals at basal (B) and after a hyperinsulinemic euglycemic clamp (Insulin (I)). Lane intensities were quantified between 15 and 300 kDa. Sample size of 8–10 people per group. Data are mean ± SEM and analyzed by two-way repeated measures ANOVA; ** indicates p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay

    Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Loss of skeletal muscle OGT alters global muscle metabolism. (A) Metabolome heat map of gastrocnemius muscle from clamped mice. Metabolites were measured using LC-MS. (B) Glycolytic metabolites from metabolome. Fructose 1 6-bisphosphate (Fru1,6P) and phosphoenolpyruvate (PEP). (C) Pyruvate dehydrogenase (PDH) activity in the gastrocnemius muscle was assayed with the substrate [1– 14 C] pyruvate by measuring enzyme-catalyzed release of 14 CO2. (D) Immunoblot of PDH phosphorylation at pS293 and pS300 in gastrocnemius from clamped mice. (E) Illustration of metabolites and regulation of glycolysis and hexosamine biosynthetic pathway. (F) UDP-GlcNAc levels from the metabolomics analysis. (G) Immunoblot of GFAT1 in gastrocnemius from clamped mice. (H) Acetylcarnitine levels from the metabolomics analysis. (I) Pantothenic acid levels from the metabolomics analysis. (J) Immunoblot of HSL and HSL pS563 in gastrocnemius from clamped mice. (K) Immunoblot of ATGL in gastrocnemius from clamped mice. (L) Complete oxidation of [1– 14 C]-palmitic acid to CO 2 in gastrocnemius muscle of WT and mKO mice. (M) Incomplete oxidation of [1– 14 C]-palmitic acid to acid-soluble metabolites (ASM) in gastrocnemius muscle. Data represent means ± SEM from n = 8–10, 18-week-old (Panel A, B, C, F, G, H, I, J, and K) and n = 10, 16-week-old (Panel D, L, and M) male mice in each genotype. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

    Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Journal: Molecular Metabolism

    Article Title: Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

    doi: 10.1016/j.molmet.2018.02.010

    Figure Lengend Snippet: Lack of OGT in skeletal muscle affects glucose homeostasis and insulin signaling. (A – B) Blood glucose levels before and during a hyperinsulinemic euglycemic clamp performed using 4 mU/kg/min of insulin. (C – D) Glucose infusion rates (GIR) during clamp. (E) Plasma insulin levels before and during steady state of the clamp. (F) Glucose uptake in skeletal muscles after steady state. (G) Glucose uptake in adipose tissues after steady state. (H) Glucose uptake in BAT, heart, and brain after steady state. (I) Glycogen in gastrocnemius, triceps, quadriceps and liver from clamped mice. (J) Glycogen in gastrocnemius and liver from 5-hour fasted mice. (K) Immunoblot of Akt and Akt phosphorylation of pS473 and pT308 in gastrocnemius. (L) Immunoblot of Akt1 in gastrocnemius from clamped mice. (M) Immunoblot of TBC1D4 and TBC1D4 phosphorylation of, pS324, pT642, and pS711 in gastrocnemius. (N) Immunoblot of GLUT4 in gastrocnemius. Data represent means ± SEM from n = 8–10 mice in each genotype. Mice were 18-week-old males. *p

    Article Snippet: Antibodies used for Western blot analyses were: anti-OGT (#SAB2101676 Sigma), anti-O-GlcNAc (#2739 Abcam), anti-ATP synthase β subunit (#A-21351 ThermoFisher Scientific), anti-succinate dehydrogenase A subunit (SDHA) (#14715 Abcam), OGT (#5368 CST), O-GlcNAc (CTD110.6 #9875 CST), OGA (#124807 Abcam), Hexokinase II (#2867 CST), PFK1 (#166722 Santa Cruz Biotechnology (SCBT)), PDH (#C54G1 CST), PDH pS293 (#ABS204 Millipore), PDH pS300 (#ABS194 Millipore), ACC pS79/257 (#07–300 Millipore), Insulin receptor β (#3025 CST), IRS1 (#23822 CST), AKT 1/2/3 (#9272 CST), AKT pT308 (#9275 CST), AKT pS473 (#9271 CST), AKT1 (#5298 SCBT), TBC1D4 (#07–741 Millipore), TBC1D4 pS711 (Custom), TBC1D4 pS324 (Custom), ATGL (#2138 CST), HSL (#4107 CST), HSL pS563 (#4139 CST), GFAT1 (#125069 Abcam), GLUT4 (#PA1-1065 Thermo Fisher).

    Techniques: Mouse Assay