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Abcam uk rabbit anti glucose 6 phosphatase
Upregulation of FGF21 production by GLP-1 analogs is recapitulated in isolated human hepatocytes and in patients with type 2 diabetes (T2D). (a-d) Human primary hepatocytes were cultured with or without liraglutide (Lira, 100 nM) for 24 h. Intracellular (a) and supernatant (b) levels of FGF21 protein were examined. The protein levels of <t>glucose</t> 6 phosphatase (G6Pase) (c) and phosphoenolpyruvate carboxykinase (PEPCK) (d) were detected. Lines with each color represent different batches (a). Data in b-d are shown as means ± S.D. Statistical analysis was carried out using unpaired two-tailed Student's t- test. ⁎ P
Uk Rabbit Anti Glucose 6 Phosphatase, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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93/100 stars

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1) Product Images from "Liver-derived fibroblast growth factor 21 mediates effects of glucagon-like peptide-1 in attenuating hepatic glucose output"

Article Title: Liver-derived fibroblast growth factor 21 mediates effects of glucagon-like peptide-1 in attenuating hepatic glucose output

Journal: EBioMedicine

doi: 10.1016/j.ebiom.2019.02.037

Upregulation of FGF21 production by GLP-1 analogs is recapitulated in isolated human hepatocytes and in patients with type 2 diabetes (T2D). (a-d) Human primary hepatocytes were cultured with or without liraglutide (Lira, 100 nM) for 24 h. Intracellular (a) and supernatant (b) levels of FGF21 protein were examined. The protein levels of glucose 6 phosphatase (G6Pase) (c) and phosphoenolpyruvate carboxykinase (PEPCK) (d) were detected. Lines with each color represent different batches (a). Data in b-d are shown as means ± S.D. Statistical analysis was carried out using unpaired two-tailed Student's t- test. ⁎ P
Figure Legend Snippet: Upregulation of FGF21 production by GLP-1 analogs is recapitulated in isolated human hepatocytes and in patients with type 2 diabetes (T2D). (a-d) Human primary hepatocytes were cultured with or without liraglutide (Lira, 100 nM) for 24 h. Intracellular (a) and supernatant (b) levels of FGF21 protein were examined. The protein levels of glucose 6 phosphatase (G6Pase) (c) and phosphoenolpyruvate carboxykinase (PEPCK) (d) were detected. Lines with each color represent different batches (a). Data in b-d are shown as means ± S.D. Statistical analysis was carried out using unpaired two-tailed Student's t- test. ⁎ P

Techniques Used: Isolation, Cell Culture, Two Tailed Test

FGF21 is a key mediator of inhibition of gluconeogenesis by GLP-1 analogs in cultured hepatocytes. (a-d) Effect of FGF21 neutralizing antibody (Ab, 5 μg/mL) on glucose 6 phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK) protein levels in mouse primary hepatocytes incubated with exendin-4 (Ex) (a,b) or liraglutide (Lira) (c,d). (e-h) HepG2 cells were transfected with FGF21 siRNA (si-FGF21) for 48 h and then cultured with Ex (e,f) or Lira (g,h) for an additional 24 h. The mRNA (upper panel) and protein (lower panel) levels of G6Pase and PEPCK were detected. (i-l) Primary hepatocytes isolated from Fgf21 knockout (KO) and wild-type (WT) mice were cultured with Ex (i,j) or Lira (k,l). The mRNA (upper panel) and protein (lower panel) levels of G6Pase and PEPCK were measured. Data are shown as means ± S.D. n = 4. One-way ANOVA, followed by the post hoc Tukey-Kramer test, was used for statistical analysis. Data in a-h, ⁎ P
Figure Legend Snippet: FGF21 is a key mediator of inhibition of gluconeogenesis by GLP-1 analogs in cultured hepatocytes. (a-d) Effect of FGF21 neutralizing antibody (Ab, 5 μg/mL) on glucose 6 phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK) protein levels in mouse primary hepatocytes incubated with exendin-4 (Ex) (a,b) or liraglutide (Lira) (c,d). (e-h) HepG2 cells were transfected with FGF21 siRNA (si-FGF21) for 48 h and then cultured with Ex (e,f) or Lira (g,h) for an additional 24 h. The mRNA (upper panel) and protein (lower panel) levels of G6Pase and PEPCK were detected. (i-l) Primary hepatocytes isolated from Fgf21 knockout (KO) and wild-type (WT) mice were cultured with Ex (i,j) or Lira (k,l). The mRNA (upper panel) and protein (lower panel) levels of G6Pase and PEPCK were measured. Data are shown as means ± S.D. n = 4. One-way ANOVA, followed by the post hoc Tukey-Kramer test, was used for statistical analysis. Data in a-h, ⁎ P

Techniques Used: Inhibition, Cell Culture, Incubation, Transfection, Isolation, Knock-Out, Mouse Assay

FGF21 is a key mediator of inhibition of glucose output by GLP-1 analogs in the liver of diabetic mice. (a-f) Eight-week-old male db/db mice were treated with exenatide (Ex) or PBS for 2 weeks, and half of the Ex-treated mice were given a single intraperitoneal injection of 8 μg FGF21 neutralizing antibody (Ab) before being subjected to the intraperitoneal glucose tolerance test (IPGTT), insulin tolerance test (ITT) and pyruvate tolerance test (PTT). Blood glucose levels (a) and their areas under curve (AUC) (b), and plasma insulin levels (c) during the IPGTT are shown. Blood glucose levels during ITT (d) and PTT (e), and the AUC of blood glucose during PTT (f) are shown. Age-matched male db/m mice treated with PBS were included as a normal control. n = 8 per group. (g-l) The mRNA (g,h), protein (i,j) levels and activity (k,l) of glucose 6 phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK) in the liver tissues of db/db mice were measured (n = 8). (m-p) Thirteen-week-old male diabetic Pax6 m/+ mice were treated for 2 weeks with liraglutide (Lira, 0.2 mg/kg) or PBS via subcutaneous injection twice daily. Age-matched male C57BL/6 mice treated with PBS were used as a normal control. The protein levels (m,n) and activity (o,p) of G6Pase and PEPCK in the liver tissues were measured (n = 3). Data are shown as means ± S.D. One-way ANOVA, followed by the post hoc Tukey-Kramer test, was used for statistical analysis. Data in a-l, ⁎ P
Figure Legend Snippet: FGF21 is a key mediator of inhibition of glucose output by GLP-1 analogs in the liver of diabetic mice. (a-f) Eight-week-old male db/db mice were treated with exenatide (Ex) or PBS for 2 weeks, and half of the Ex-treated mice were given a single intraperitoneal injection of 8 μg FGF21 neutralizing antibody (Ab) before being subjected to the intraperitoneal glucose tolerance test (IPGTT), insulin tolerance test (ITT) and pyruvate tolerance test (PTT). Blood glucose levels (a) and their areas under curve (AUC) (b), and plasma insulin levels (c) during the IPGTT are shown. Blood glucose levels during ITT (d) and PTT (e), and the AUC of blood glucose during PTT (f) are shown. Age-matched male db/m mice treated with PBS were included as a normal control. n = 8 per group. (g-l) The mRNA (g,h), protein (i,j) levels and activity (k,l) of glucose 6 phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK) in the liver tissues of db/db mice were measured (n = 8). (m-p) Thirteen-week-old male diabetic Pax6 m/+ mice were treated for 2 weeks with liraglutide (Lira, 0.2 mg/kg) or PBS via subcutaneous injection twice daily. Age-matched male C57BL/6 mice treated with PBS were used as a normal control. The protein levels (m,n) and activity (o,p) of G6Pase and PEPCK in the liver tissues were measured (n = 3). Data are shown as means ± S.D. One-way ANOVA, followed by the post hoc Tukey-Kramer test, was used for statistical analysis. Data in a-l, ⁎ P

Techniques Used: Inhibition, Mouse Assay, Injection, Activity Assay

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    Abcam glucose 6 phosphatase
    Loss of Argonaute2 in liver reduces hepatic gluconeogenesis. A , Western blotting analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls (n = 3). B , qRT-PCR analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 7). C , Random and fasting glucose measurements in 12-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls (n = 8–11). D , Body weight in male Alb -Cre, Ago2 flox/flox mice (n = 13) and littermate controls (n = 14) from 4 to 12 weeks of age. E , Glucose measurements during an insulin tolerance test on 10-week old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 6). F , Glucose measurements during a glucose tolerance test on 11-week old Alb -Cre, Ago2 flox/flox mice (n = 7) and littermate controls (n = 6). G , Glucose measurements during a pyruvate tolerance test on 12-week old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 6). H , Glucose measurements and body weight in the Alb -Cre, Ago2 flox/flox mice (n = 8) and littermate controls (n = 4) on normal chow (Chow) or ketogenic diet (Keto, n = 8) for 30 days. I , qRT-PCR analysis of G6Pase , Pepck and Pyruvate carboxylase ( Pc1 ) from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls after overnight fasting (n = 6). J , Western blotting analysis of <t>glucose-6</t> phosphatase (G6Pase), glucokinase (Gck) and glycogen synthase (Gly synth) from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls after overnight fasting (n = 3). K , Plasma insulin measurements in 12-week old Alb -Cre, Ago2 flox/flox mice (n = 7) and littermate controls after overnight fasting (n = 6). L , Western blotting analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls during random-fed and overnight fasted states (16 h) (n = 2). M , Western blotting analysis of insulin receptor (InsR), phosphorylated Akt2 at Ser473 (p-Akt2), Akt2, phosphorylated FoxO1 at Thr24 (p-FoxO1) and FoxO1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls during random-fed and overnight fasted states (16 h) (n = 2). Results are presented as mean ± SEM. * P
    Glucose 6 Phosphatase, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glucose 6 phosphatase/product/Abcam
    Average 93 stars, based on 1 article reviews
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    glucose 6 phosphatase - by Bioz Stars, 2022-01
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    Loss of Argonaute2 in liver reduces hepatic gluconeogenesis. A , Western blotting analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls (n = 3). B , qRT-PCR analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 7). C , Random and fasting glucose measurements in 12-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls (n = 8–11). D , Body weight in male Alb -Cre, Ago2 flox/flox mice (n = 13) and littermate controls (n = 14) from 4 to 12 weeks of age. E , Glucose measurements during an insulin tolerance test on 10-week old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 6). F , Glucose measurements during a glucose tolerance test on 11-week old Alb -Cre, Ago2 flox/flox mice (n = 7) and littermate controls (n = 6). G , Glucose measurements during a pyruvate tolerance test on 12-week old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 6). H , Glucose measurements and body weight in the Alb -Cre, Ago2 flox/flox mice (n = 8) and littermate controls (n = 4) on normal chow (Chow) or ketogenic diet (Keto, n = 8) for 30 days. I , qRT-PCR analysis of G6Pase , Pepck and Pyruvate carboxylase ( Pc1 ) from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls after overnight fasting (n = 6). J , Western blotting analysis of glucose-6 phosphatase (G6Pase), glucokinase (Gck) and glycogen synthase (Gly synth) from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls after overnight fasting (n = 3). K , Plasma insulin measurements in 12-week old Alb -Cre, Ago2 flox/flox mice (n = 7) and littermate controls after overnight fasting (n = 6). L , Western blotting analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls during random-fed and overnight fasted states (16 h) (n = 2). M , Western blotting analysis of insulin receptor (InsR), phosphorylated Akt2 at Ser473 (p-Akt2), Akt2, phosphorylated FoxO1 at Thr24 (p-FoxO1) and FoxO1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls during random-fed and overnight fasted states (16 h) (n = 2). Results are presented as mean ± SEM. * P

    Journal: Molecular Metabolism

    Article Title: Control of hepatic gluconeogenesis by Argonaute2

    doi: 10.1016/j.molmet.2018.10.003

    Figure Lengend Snippet: Loss of Argonaute2 in liver reduces hepatic gluconeogenesis. A , Western blotting analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls (n = 3). B , qRT-PCR analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 7). C , Random and fasting glucose measurements in 12-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls (n = 8–11). D , Body weight in male Alb -Cre, Ago2 flox/flox mice (n = 13) and littermate controls (n = 14) from 4 to 12 weeks of age. E , Glucose measurements during an insulin tolerance test on 10-week old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 6). F , Glucose measurements during a glucose tolerance test on 11-week old Alb -Cre, Ago2 flox/flox mice (n = 7) and littermate controls (n = 6). G , Glucose measurements during a pyruvate tolerance test on 12-week old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls (n = 6). H , Glucose measurements and body weight in the Alb -Cre, Ago2 flox/flox mice (n = 8) and littermate controls (n = 4) on normal chow (Chow) or ketogenic diet (Keto, n = 8) for 30 days. I , qRT-PCR analysis of G6Pase , Pepck and Pyruvate carboxylase ( Pc1 ) from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice (n = 5) and littermate controls after overnight fasting (n = 6). J , Western blotting analysis of glucose-6 phosphatase (G6Pase), glucokinase (Gck) and glycogen synthase (Gly synth) from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls after overnight fasting (n = 3). K , Plasma insulin measurements in 12-week old Alb -Cre, Ago2 flox/flox mice (n = 7) and littermate controls after overnight fasting (n = 6). L , Western blotting analysis of Ago2 and Ago1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls during random-fed and overnight fasted states (16 h) (n = 2). M , Western blotting analysis of insulin receptor (InsR), phosphorylated Akt2 at Ser473 (p-Akt2), Akt2, phosphorylated FoxO1 at Thr24 (p-FoxO1) and FoxO1 from liver of 8-week-old Alb -Cre, Ago2 flox/flox mice and littermate controls during random-fed and overnight fasted states (16 h) (n = 2). Results are presented as mean ± SEM. * P

    Article Snippet: 2.2.2 Antibodies for western blot analysis The following primary antibodies were used for western blotting at 1:1000 dilution: Ago2 (MBL CM004-3), Ago1 (MBL RN028PW), Glucokinase (Abcam ab88056), β-Actin (Sigma A1978), GAPDH (Abcam ab8245), α-Tubulin (Sigma, T6557), Glucose-6-phosphatase (Abcam ab83690), Glycogen synthase (Cell Signaling, #3893), Insulin receptor (Cell Signaling, #3025), Akt2 (Cell Signaling, #2964), phospho-Akt2 (Ser474) (Cell Signaling, #8599), FoxO1 (Cell Signaling, #2880), phospho-FoxO1 (Cell Signaling, #9464), Acetyl-CoA Carboxylase (Cell Signaling, #3676), phospho-Acetyl-CoA Carboxylase (Cell Signaling, #11818), Fatty Acid Synthase (Cell Signaling, #3180), AMPKα (Cell Signaling, #2532), and phospho-AMPKα (Cell Signaling, #2535).

    Techniques: Western Blot, Mouse Assay, Quantitative RT-PCR

    Effect of STZ treatment on enzymatic pathway of glucose degradation. Catalytic activities of hexose-kinase (HK, Panel A ), glucose-6-phosphatase (G6Pase, Panel B ), Phosphofructokinase (PFK, Panel C ), glucose-6-phosphate dehydrogenase (G6PD, Panel D ), and hexose-6-phosphate dehydrogenase (H6PD, Panel E ) in anterior (Ant) and posterior (Post) brain areas in control ( n = 6 for each area, blue) and STZ mice ( n = 6 for each area, brown). STZ treatment significantly reduced H6PD catalytic function in posterior brain areas, while it did not affect the main enzymes involved in glucose entrapment, release, and cytosolic catabolism. Data are expressed as mean ± SD.

    Journal: International Journal of Molecular Sciences

    Article Title: 18F-Fluorodeoxyglucose Positron Emission Tomography Tracks the Heterogeneous Brain Susceptibility to the Hyperglycemia-Related Redox Stress

    doi: 10.3390/ijms21218154

    Figure Lengend Snippet: Effect of STZ treatment on enzymatic pathway of glucose degradation. Catalytic activities of hexose-kinase (HK, Panel A ), glucose-6-phosphatase (G6Pase, Panel B ), Phosphofructokinase (PFK, Panel C ), glucose-6-phosphate dehydrogenase (G6PD, Panel D ), and hexose-6-phosphate dehydrogenase (H6PD, Panel E ) in anterior (Ant) and posterior (Post) brain areas in control ( n = 6 for each area, blue) and STZ mice ( n = 6 for each area, brown). STZ treatment significantly reduced H6PD catalytic function in posterior brain areas, while it did not affect the main enzymes involved in glucose entrapment, release, and cytosolic catabolism. Data are expressed as mean ± SD.

    Article Snippet: The membrane was blocked for 1 h with Tris Buffered Saline (TBS) plus 0.15% Tween 20 (TBSt) containing 5% nonfat dry milk and incubated overnight at 4 °C with the following rabbit polyclonal antibodies: anti-HK II (1:1000, Cell Signalling #2867), anti-G6Pase (1:1000, Abcam, ab93857), anti-PFKP (1:200, Cell Signalling #8164), anti-G6PD (1:1000, Abcam, ab210702), anti_H6PD (1:1000, Abcam, ab170895), or anti-actin (1:10,000, Thermo-Fisher MA5-11869).

    Techniques: Mouse Assay

    Mitochondrial morphology is deranged in the livers of G6PC KO mice and G6PC KD AML-12 cells (96 hour knockdown). Mitochondrial morphology was analysed in ultrathin sections of mouse liver ( A ) and AML-12 cells ( B ) by electron microscopy. Mitochondrial morphology in the wild-type mice and control cells were within normal limits, whereas in the KO mice and KD cells, the mitochondria were distended, and swollen, with effacement of the cristae, disruption of the mitochondrial membranes, and influx of cytoplasmic contents into the mitochondria. Representative images from the mouse liver ( A ) and KD cells ( B ) are displayed. Scale bars are marked on each image. Frames box areas shown at high magnification.

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: Mitochondrial morphology is deranged in the livers of G6PC KO mice and G6PC KD AML-12 cells (96 hour knockdown). Mitochondrial morphology was analysed in ultrathin sections of mouse liver ( A ) and AML-12 cells ( B ) by electron microscopy. Mitochondrial morphology in the wild-type mice and control cells were within normal limits, whereas in the KO mice and KD cells, the mitochondria were distended, and swollen, with effacement of the cristae, disruption of the mitochondrial membranes, and influx of cytoplasmic contents into the mitochondria. Representative images from the mouse liver ( A ) and KD cells ( B ) are displayed. Scale bars are marked on each image. Frames box areas shown at high magnification.

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques: Mouse Assay, Electron Microscopy

    The tri-carboxylic acid cycle function is impaired in G6PC KO mice. ( A ) Organic acid analysis in the livers G6PC KO mice (KO, n = 6) compared to wild-type (WT, n = 10) mice. ( B ) Analysis of amino acid levels in the same samples. ( C ) Schematic of the changes of the species in the TCA cycle. Species in green were increased in G6PC KO mouse livers relative to WT livers, while those in red were decreased. Species in blue showed no significant change between groups, while those in black were not directly measured. For all parts, error bars represent SEM, *represents p

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: The tri-carboxylic acid cycle function is impaired in G6PC KO mice. ( A ) Organic acid analysis in the livers G6PC KO mice (KO, n = 6) compared to wild-type (WT, n = 10) mice. ( B ) Analysis of amino acid levels in the same samples. ( C ) Schematic of the changes of the species in the TCA cycle. Species in green were increased in G6PC KO mouse livers relative to WT livers, while those in red were decreased. Species in blue showed no significant change between groups, while those in black were not directly measured. For all parts, error bars represent SEM, *represents p

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques: Mouse Assay

    Key factors involved in mitochondrial biogenesis are reduced following G6PC KD for 96 hours in AML-12 cells. ( A ) Analysis of mRNA expression of genes involved in mitochondrial biogenesis, quality control and mitophagy shows a decrease in expression of genes related to mitochondrial biogenesis, with no change in expression of key fission and fusion genes. (n = 3). ( B ) Protein levels of TFAM are reduced following G6PC KD, however those of NRF1 show no significant change (n = 5). *Represents p

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: Key factors involved in mitochondrial biogenesis are reduced following G6PC KD for 96 hours in AML-12 cells. ( A ) Analysis of mRNA expression of genes involved in mitochondrial biogenesis, quality control and mitophagy shows a decrease in expression of genes related to mitochondrial biogenesis, with no change in expression of key fission and fusion genes. (n = 3). ( B ) Protein levels of TFAM are reduced following G6PC KD, however those of NRF1 show no significant change (n = 5). *Represents p

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques: Expressing

    Mitochondrial content is reduced in GSDIa models. ( A ) The protein levels of various key mitochondrial proteins are reduced following G6PC KD for 96 hours in AML-12 cells (n = 3). ( B ) The copy number of mitochondrial DNA is also reduced following G6PC KD for 96 hours in AML-12 cells (n = 6). ( C ) G6PC KO mouse livers also show decreased levels of mitochondrial proteins (n = 3). *Represents p

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: Mitochondrial content is reduced in GSDIa models. ( A ) The protein levels of various key mitochondrial proteins are reduced following G6PC KD for 96 hours in AML-12 cells (n = 3). ( B ) The copy number of mitochondrial DNA is also reduced following G6PC KD for 96 hours in AML-12 cells (n = 6). ( C ) G6PC KO mouse livers also show decreased levels of mitochondrial proteins (n = 3). *Represents p

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques:

    Protein levels of key transcription factors involved in mitochondrial biogenesis. ( A ) PGC1α levels are reduced in G6PC KD AML-12 cells (n = 5). ( B ) PGC1α levels are also reduced in G6PC KO mouse livers (n = 3). ( C ) Levels of ERRα are unchanged in G6PC KO mouse livers as compared to WT livers (n = 3). *Represents p

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: Protein levels of key transcription factors involved in mitochondrial biogenesis. ( A ) PGC1α levels are reduced in G6PC KD AML-12 cells (n = 5). ( B ) PGC1α levels are also reduced in G6PC KO mouse livers (n = 3). ( C ) Levels of ERRα are unchanged in G6PC KO mouse livers as compared to WT livers (n = 3). *Represents p

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques:

    Mitochondrial respiration is impaired in G6PC KD versus control siRNA treated AML-12 cells. G6PC was knocked-down in AML-12 cells for 96 hours, then mitochondrial oximetry analysis was performed using a Seahorse XF24 mitochondrial flux analyser. Basal respiration, ATP turnover, Maximal Respiration, and Spare Mitochondrial capacity were measured/calculated as described in the methods section. Oxygen consumption was normalised to total cellular protein content. For all parts, n = 6, error bars represent SEM, *represents p

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: Mitochondrial respiration is impaired in G6PC KD versus control siRNA treated AML-12 cells. G6PC was knocked-down in AML-12 cells for 96 hours, then mitochondrial oximetry analysis was performed using a Seahorse XF24 mitochondrial flux analyser. Basal respiration, ATP turnover, Maximal Respiration, and Spare Mitochondrial capacity were measured/calculated as described in the methods section. Oxygen consumption was normalised to total cellular protein content. For all parts, n = 6, error bars represent SEM, *represents p

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques:

    The mitochondrial apoptosis pathway is up-regulated in GSD1a. ( A ) Mitochondrial membrane potential is reduced in G6PC KD AML-12 cells 72 hours after knockdown. ( B ) Cytosolic cytochrome c is increased in G6PC KD AML-12 cells. ( C ) Caspase 9 and caspase 3 cleavage is increased in G6PC KD AML-12 cells. ( D , E ) Caspase 9 ( D ) and caspase 3 ( E ) cleavage is increasted in G6PC KO mice. For all parts except A, n = 3, error bars represent SEM, and *represents p

    Journal: Scientific Reports

    Article Title: Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa)

    doi: 10.1038/srep44408

    Figure Lengend Snippet: The mitochondrial apoptosis pathway is up-regulated in GSD1a. ( A ) Mitochondrial membrane potential is reduced in G6PC KD AML-12 cells 72 hours after knockdown. ( B ) Cytosolic cytochrome c is increased in G6PC KD AML-12 cells. ( C ) Caspase 9 and caspase 3 cleavage is increased in G6PC KD AML-12 cells. ( D , E ) Caspase 9 ( D ) and caspase 3 ( E ) cleavage is increasted in G6PC KO mice. For all parts except A, n = 3, error bars represent SEM, and *represents p

    Article Snippet: The antibody recognizing G6PC (83690) came from Abcam.

    Techniques: Mouse Assay

    FBP1, but not PFKL, expression decreases in ccRCC tumours and correlates with tumour stages and patient prognosis IHC staining of the kidney tissue microarray as shown in Fig. 1( b ) with G6PC ( a ), PCK1 ( b ), and PFKL ( c ) antibodies. T: ccRCC tumour; N: adjacent normal kidney. Quantification of each staining is shown on the right. d , Normalized RNASeq reads of PFKL in 69 normal kidneys and 480 ccRCC tumours grouped into Stage I-IV by TCGA. e , Kaplan-Meier survival curve of 429 ccRCC patients enrolled in the TCGA database. Patients were equally divided into two groups (top and bottom 50% PFKL expression) based on PFKL expression levels in their tumours.

    Journal: Nature

    Article Title: Fructose-1, 6-bisphosphatase opposes renal carcinoma progression

    doi: 10.1038/nature13557

    Figure Lengend Snippet: FBP1, but not PFKL, expression decreases in ccRCC tumours and correlates with tumour stages and patient prognosis IHC staining of the kidney tissue microarray as shown in Fig. 1( b ) with G6PC ( a ), PCK1 ( b ), and PFKL ( c ) antibodies. T: ccRCC tumour; N: adjacent normal kidney. Quantification of each staining is shown on the right. d , Normalized RNASeq reads of PFKL in 69 normal kidneys and 480 ccRCC tumours grouped into Stage I-IV by TCGA. e , Kaplan-Meier survival curve of 429 ccRCC patients enrolled in the TCGA database. Patients were equally divided into two groups (top and bottom 50% PFKL expression) based on PFKL expression levels in their tumours.

    Article Snippet: Antibodies detecting HIF1α (NB100-134) and HIF2α (NB100-122) were obtained from Novus, and antibodies against PCK1 (ab28455), G6PC (ab83690), and PFKL (ab37583) were from Abcam.

    Techniques: Expressing, Immunohistochemistry, Staining, Microarray

    Pan-metabolomic analysis of ccRCC tumour and adjacent normal kidney tissues a , Heatmap showing the relative concentration of 418 metabolites detected in 20 primary ccRCC tumours and adjacent normal kidneys. Metabolites were extracted from frozen tissue samples and analysed by the Thermo-Finnigan GC-MS and LC-MS/MS systems. Raw data of each metabolite was rescaled to set the median equal to 1. All metabolites were clustered according to their related pathways (based on KEGG) and then plotted as a heatmap. b , Metabolic genes involved in “carbohydrate storage” differentially expressed in ccRCC tumour vs. normal tissue. G6PC, glucose-6-phosphatase, catalytic subunit; PCK1, phosphoenolpyruvate carboxykinase 1; FBP1, fructose-1, 6-bisphosphatase 1. c , Illustration of central carbon metabolism, including glycolysis, gluconeogenesis, pentose phosphate pathway, and the TCA cycle. Enzymes controlling glycolysis (HK, hexokinase; PFK, phosphofructokinase; PKM, pyruvate kinase type M) are highlighted in red, while enzymes controlling gluconeogenesis (G6P, glucose-6-phosphatase; FBP, fructose-1, 6-bisphosphatase; PCK, phosphoenolpyruvate carboxykinase) are highlighted in green. G6P, glucose 6-phosphate; F6P, fructose 6-phosphate; F-1, 6-BP, fructose 1, 6-bisphosphate; DHAP, dihydroxyacetone phosphate; GAP, glyceraldehyde 3-phosphate; R5P, ribose 5-phosphate; X5P, xylulose 5-phosphate; E4P, erythrose 4-phosphate; S7P, sedoheptulose 7-phosphate; PEP, phosphoenolpyruvate; Pyr, pyruvate; Ac-CoA, acetyl-CoA; Lac, lactate; Cit, citrate; αKG, alpha-ketoglutarate; Glu, glutamate; Suc, succinate; Fum, fumarate; Mal, malate; Oac, oxaloacetate; Asp, aspartate; G-SH, reduced glutathione.

    Journal: Nature

    Article Title: Fructose-1, 6-bisphosphatase opposes renal carcinoma progression

    doi: 10.1038/nature13557

    Figure Lengend Snippet: Pan-metabolomic analysis of ccRCC tumour and adjacent normal kidney tissues a , Heatmap showing the relative concentration of 418 metabolites detected in 20 primary ccRCC tumours and adjacent normal kidneys. Metabolites were extracted from frozen tissue samples and analysed by the Thermo-Finnigan GC-MS and LC-MS/MS systems. Raw data of each metabolite was rescaled to set the median equal to 1. All metabolites were clustered according to their related pathways (based on KEGG) and then plotted as a heatmap. b , Metabolic genes involved in “carbohydrate storage” differentially expressed in ccRCC tumour vs. normal tissue. G6PC, glucose-6-phosphatase, catalytic subunit; PCK1, phosphoenolpyruvate carboxykinase 1; FBP1, fructose-1, 6-bisphosphatase 1. c , Illustration of central carbon metabolism, including glycolysis, gluconeogenesis, pentose phosphate pathway, and the TCA cycle. Enzymes controlling glycolysis (HK, hexokinase; PFK, phosphofructokinase; PKM, pyruvate kinase type M) are highlighted in red, while enzymes controlling gluconeogenesis (G6P, glucose-6-phosphatase; FBP, fructose-1, 6-bisphosphatase; PCK, phosphoenolpyruvate carboxykinase) are highlighted in green. G6P, glucose 6-phosphate; F6P, fructose 6-phosphate; F-1, 6-BP, fructose 1, 6-bisphosphate; DHAP, dihydroxyacetone phosphate; GAP, glyceraldehyde 3-phosphate; R5P, ribose 5-phosphate; X5P, xylulose 5-phosphate; E4P, erythrose 4-phosphate; S7P, sedoheptulose 7-phosphate; PEP, phosphoenolpyruvate; Pyr, pyruvate; Ac-CoA, acetyl-CoA; Lac, lactate; Cit, citrate; αKG, alpha-ketoglutarate; Glu, glutamate; Suc, succinate; Fum, fumarate; Mal, malate; Oac, oxaloacetate; Asp, aspartate; G-SH, reduced glutathione.

    Article Snippet: Antibodies detecting HIF1α (NB100-134) and HIF2α (NB100-122) were obtained from Novus, and antibodies against PCK1 (ab28455), G6PC (ab83690), and PFKL (ab37583) were from Abcam.

    Techniques: Concentration Assay, Gas Chromatography-Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    FBP1 inhibits HIF and pseudohypoxia in ccRCC tumour cells a , Western blot analysis of indicated proteins in HK-2, RCC4, and RCC10 cells. Oxygen consumption in RCC4 ( b ) and RCC10 ( c ) cells expressing vector or FBP1 was measured using the MitoXpress dye as described in Methods . Antimycin A (an inhibitor of mitochondrial respiration) was used as negative control. d , Carbon fate map showing the isotopomer distribution of indicated metabolites derived from [U- 13 C] glutamine. Filled circles indicate 13 C carbons derived from [U- 13 C] glutamine, whereas open circles represent carbons derived from endogenous sources. Note that by the PCK or ME pathway, M4 malate generates M3 pyruvate, which re-enters the TCA cycle through the PDH flux (coloured in red) to produce M6 or M2 citrate. ME, malic enzyme; PDH, pyruvate dehydrogenase. e , M1-M6 isotopomer distribution of citrate in RCC10 cells expressing vector or FBP1, labelled with [U- 13 C] glutamine. f , The enrichment ratio of M6 or M2 citrate to M3 pyruvate in RCC10 cells expressing vector or FBP1, labelled with [U- 13 C] glutamine. Note that this ratio is an indication of PDH activity. g , HIF reporter activity in RCC10 cells transfected with vector or V5-tagged G6PC. Protein levels of expressed V5-G6PC are shown on the right. h , 480 ccRCC tumours from TCGA database were equally divided into two groups (top and bottom 50% G6PC expression) based on G6PC expression levels, and their relative HIF activities were quantified and plotted as described in Methods. N.S., not significant. qRT-PCR analysis of HIF target genes in RCC4 ( i ), hypoxic A549 ( j ), or normoxic RCC10VHL ( k ) cells ectopically expressing vector or FBP1. Experiments were repeated twice. Values represent mean±s.d. (technical triplicates from a representative experiment) *p

    Journal: Nature

    Article Title: Fructose-1, 6-bisphosphatase opposes renal carcinoma progression

    doi: 10.1038/nature13557

    Figure Lengend Snippet: FBP1 inhibits HIF and pseudohypoxia in ccRCC tumour cells a , Western blot analysis of indicated proteins in HK-2, RCC4, and RCC10 cells. Oxygen consumption in RCC4 ( b ) and RCC10 ( c ) cells expressing vector or FBP1 was measured using the MitoXpress dye as described in Methods . Antimycin A (an inhibitor of mitochondrial respiration) was used as negative control. d , Carbon fate map showing the isotopomer distribution of indicated metabolites derived from [U- 13 C] glutamine. Filled circles indicate 13 C carbons derived from [U- 13 C] glutamine, whereas open circles represent carbons derived from endogenous sources. Note that by the PCK or ME pathway, M4 malate generates M3 pyruvate, which re-enters the TCA cycle through the PDH flux (coloured in red) to produce M6 or M2 citrate. ME, malic enzyme; PDH, pyruvate dehydrogenase. e , M1-M6 isotopomer distribution of citrate in RCC10 cells expressing vector or FBP1, labelled with [U- 13 C] glutamine. f , The enrichment ratio of M6 or M2 citrate to M3 pyruvate in RCC10 cells expressing vector or FBP1, labelled with [U- 13 C] glutamine. Note that this ratio is an indication of PDH activity. g , HIF reporter activity in RCC10 cells transfected with vector or V5-tagged G6PC. Protein levels of expressed V5-G6PC are shown on the right. h , 480 ccRCC tumours from TCGA database were equally divided into two groups (top and bottom 50% G6PC expression) based on G6PC expression levels, and their relative HIF activities were quantified and plotted as described in Methods. N.S., not significant. qRT-PCR analysis of HIF target genes in RCC4 ( i ), hypoxic A549 ( j ), or normoxic RCC10VHL ( k ) cells ectopically expressing vector or FBP1. Experiments were repeated twice. Values represent mean±s.d. (technical triplicates from a representative experiment) *p

    Article Snippet: Antibodies detecting HIF1α (NB100-134) and HIF2α (NB100-122) were obtained from Novus, and antibodies against PCK1 (ab28455), G6PC (ab83690), and PFKL (ab37583) were from Abcam.

    Techniques: Western Blot, Expressing, Plasmid Preparation, Negative Control, Derivative Assay, Activity Assay, Transfection, Quantitative RT-PCR

    FBP1 expression affects cell proliferation a , Shown on the left is the western blot analysis of V5-tagged FBP1 and actin in 786-O cells ectopically expressing vector or V5-FBP1; On the right are protein levels of FBP1 and actin in HK-2 cells and 786-O cells with or without ectopic FBP1 expression. b , Anchorage-independent growth assay of 786-O cells expressing vector or V5-FBP1. c , Xenograft growth curves as performed in Fig. 1(f) . Low serum growth curve of RCC10 ( d ) and 769-P ( e ) cells expressing vector or V5-FBP1. Western blot analyses confirming FBP1 expression are shown on the right. f , Growth of A549 lung cancer cells under normoxia (21% O 2 ) or hypoxia (0.5% O 2 ) cultured in low serum medium (1% FBS), with or without FBP1 expression. g , Protein levels of V5-FBP1 and actin in A549 lung cancer cells as indicated in ( f ). h , Western blot analysis confirming the effect of FBP1 ablation in HK-2 cells. Growth curves of HK-2 cells with G6PC inhibition ( i ) or V5-PFKL expression ( j ), as compared to vector control cells in 1% serum medium. Western blot analyses of indicated proteins are shown on the right. All values represent mean±s.d. (four technical replicates, from two independent experiments). *p

    Journal: Nature

    Article Title: Fructose-1, 6-bisphosphatase opposes renal carcinoma progression

    doi: 10.1038/nature13557

    Figure Lengend Snippet: FBP1 expression affects cell proliferation a , Shown on the left is the western blot analysis of V5-tagged FBP1 and actin in 786-O cells ectopically expressing vector or V5-FBP1; On the right are protein levels of FBP1 and actin in HK-2 cells and 786-O cells with or without ectopic FBP1 expression. b , Anchorage-independent growth assay of 786-O cells expressing vector or V5-FBP1. c , Xenograft growth curves as performed in Fig. 1(f) . Low serum growth curve of RCC10 ( d ) and 769-P ( e ) cells expressing vector or V5-FBP1. Western blot analyses confirming FBP1 expression are shown on the right. f , Growth of A549 lung cancer cells under normoxia (21% O 2 ) or hypoxia (0.5% O 2 ) cultured in low serum medium (1% FBS), with or without FBP1 expression. g , Protein levels of V5-FBP1 and actin in A549 lung cancer cells as indicated in ( f ). h , Western blot analysis confirming the effect of FBP1 ablation in HK-2 cells. Growth curves of HK-2 cells with G6PC inhibition ( i ) or V5-PFKL expression ( j ), as compared to vector control cells in 1% serum medium. Western blot analyses of indicated proteins are shown on the right. All values represent mean±s.d. (four technical replicates, from two independent experiments). *p

    Article Snippet: Antibodies detecting HIF1α (NB100-134) and HIF2α (NB100-122) were obtained from Novus, and antibodies against PCK1 (ab28455), G6PC (ab83690), and PFKL (ab37583) were from Abcam.

    Techniques: Expressing, Western Blot, Plasmid Preparation, Growth Assay, Cell Culture, Inhibition