pck1 inhibitor  (TargetMol)

Journal: PLoS ONE

Article Title: The N -Reductive System Composed of Mitochondrial Amidoxime Reducing Component (mARC), Cytochrome b5 (CYB5B) and Cytochrome b5 Reductase (CYB5R) Is Regulated by Fasting and High Fat Diet in Mice

doi: 10.1371/journal.pone.0105371

Figure Lengend Snippet: Primers used for expression studies.

Article Snippet: For immuno-blot analysis, the following primary antibodies were used: anti-CYB5R3-antibody (HPA001566; Sigma-Aldrich, St. Louis, USA), anti-mARC2-antibody (HPA015085; Sigma-Aldrich, St. Louis, USA), anti-CYB5B-antibody (HPA007893; Sigma-Aldrich, St. Louis, USA), anti-PCK1-antibody (ab28455; Abcam, Cambridge, UK), anti-MOSC1-antibody (AP9754c; Abgent, San Diego, USA) for of HepG2 lysates or anti-MOSC1-antibody (ABIN503067; antibodies-online GmbH, Aachen, Germany) for murine liver and Hepa 1.6 lysates.

Techniques: Expressing

Journal: PLoS ONE

Article Title: The N -Reductive System Composed of Mitochondrial Amidoxime Reducing Component (mARC), Cytochrome b5 (CYB5B) and Cytochrome b5 Reductase (CYB5R) Is Regulated by Fasting and High Fat Diet in Mice

doi: 10.1371/journal.pone.0105371

Figure Lengend Snippet: Two groups of 12 C57BL/6W mice were fed with regular diet, one group were food deprived for 18 h (fasted) before sacrifice and liver collection, the second had full access to food and water (non-fasted). A Expressions of mARC1, mARC2, CYB5B, CYB5R and PCK1, determined with qRT-PCR, normalized on geometric mean of Ct values for Mcoln1 (Mucolipin-1) and Hmbs (hydroxymethylbilane synthase). Statistical significance was assessed by the U-test. p -values <0.05 were considered significant; (**) p -value <0.001; (***) p -value <0.0001. B Protein levels of mARC1, mARC2, CYB5B, CYB5R and PCK1 examined by Western Blot. Each sample consisted of equal protein amount of two individuals.

Article Snippet: For immuno-blot analysis, the following primary antibodies were used: anti-CYB5R3-antibody (HPA001566; Sigma-Aldrich, St. Louis, USA), anti-mARC2-antibody (HPA015085; Sigma-Aldrich, St. Louis, USA), anti-CYB5B-antibody (HPA007893; Sigma-Aldrich, St. Louis, USA), anti-PCK1-antibody (ab28455; Abcam, Cambridge, UK), anti-MOSC1-antibody (AP9754c; Abgent, San Diego, USA) for of HepG2 lysates or anti-MOSC1-antibody (ABIN503067; antibodies-online GmbH, Aachen, Germany) for murine liver and Hepa 1.6 lysates.

Techniques: Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Vascular and Liver Homeostasis in Juvenile Mice Require Endothelial Cyclic AMP-Dependent Protein Kinase A

doi: 10.3390/ijms231911419

Figure Lengend Snippet: Hepatic marker expression is not decreased under inhibition of endothelial PKA. ( A ), Albumin ( Alb ), Hnf4a and Hnf1b , G6pc and Pcx1 mRNA levels were analyzed in control and dnPKA iEC mice. Protein levels of albumin ( B ) and phosphoenolpyruvate carboxykinase (Pck1; ( C )) were analyzed by Western blotting. ( D ) Blots for albumin, Pck1 and GAPDH are shown. Here, 7 control (Tg(Cdh5-cre/ERT2)1Rha without floxed transgenes) and 9 dnPKA iEC mice from three different litters were analyzed. Mice (males and females) were injected with tamoxifen at the age of 30–35 days and dissected at the age of 61–84 days. Shown are means ± SD. Data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test ( A ) or two-tailed unpaired Student’s t -test ( B , C ). ns, not significant.

Article Snippet: Equal amounts of protein were loaded and separated by SDS-PAGE using a Criterion electrophoresis system (Bio-Rad Laboratories, Hercules, CA, USA), transferred onto the polyvinylidene fluoride membrane, and immunoblotted with specific antibodies: rabbit anti-albumin (Bio-Rad; #AHP1478); rabbit anti-Pck1 (Abcam; ab28455); or rabbit anti-GAPDH (#2118; Cell Signaling Technologies, Danvers, MA, USA).

Techniques: Marker, Expressing, Inhibition, Western Blot, Injection, Two Tailed Test

Journal: Scientific Reports

Article Title: Porphyromonas gingivalis induces entero-hepatic metabolic derangements with alteration of gut microbiota in a type 2 diabetes mouse model

doi: 10.1038/s41598-021-97868-2

Figure Lengend Snippet: Effects of oral Porphyromonas gingivalis ( Pg ) administration on mRNA and protein expression in the liver of db / db mice. ( a ) mRNA expression of Pck1 , G6pc , Foxo1 , Cyp7a1 , Fasn , Acaca , Cpt1 , Srebf1 , and Srebf2 . mRNA expression was determined using quantitative RT-PCR and was normalized against the expression of 18S rRNA mRNA. Target gene expression in Pg -treated mice was normalized against the target gene expression in CMC-treated mice, which is considered as 1. Treatment or control group variability was calculated as the ratio of [each amount/the mean amount in the control] in every chart. n = 20–21, ** P < 0.01, * P < 0.05 versus control. ( b ) Western blot analysis of PCK1, G6PC, and FOXO1 in liver tissues from db / db mice treated with Pg or CMC for 30 days. β-actin was used as the loading control. The bar graphs on the right show densitometric quantification of the amounts of PCK1, G6PC, and FOXO1, relative to that in the control. n = 4; * P < 0.05 versus control. ( c , d ) PCK1 ( c ) and FOXO1 ( d ) detection in liver tissues of Pg - or control-treated mice. Left column: Paraffin-embedded sections stained with hematoxylin and eosin (H&E). Right column: Immunohistochemical detection. Scale bars: 100 μm. ( e ) Comparison of the relative gene expression of proinflammatory cytokines ( Il-6 , Tnf-α , Ccl2 , and Cxcl10 ) in the liver tissues of Pg - and control-treated db / db mice. The data were normalized and analyzed as described for ( a ). n = 20–21. Pck1, Phosphoenolpyruvate carboxykinase 1; G6pc, Glucose-6-phosphatase; Foxo1, Forkhead box protein O1; Cpt1c, Carnitine palmitoyltransferase 1c; Fasn, Fatty acid synthase; Acaca, Acetyl-Coenzyme A carboxylase alpha; Srebf1, Sterol regulatory element-binding transcription factor 1; Srebf2, Sterol regulatory element-binding transcription factor 2; Il-6, Interleukin 6; Tnf-α, Tumor necrosis factor-α; Ccl2, Chemokine (C–C motif) ligand 2 ; Cxcl10, C-X-C motif chemokine ligand 10.

Article Snippet: Immunoblotting was performed using the following primary antibodies: PCK1 (1:1000; ab28455, Abcam, Toronto, Canada), G6PC (ab83690; Abcam), FOXO1 (1:000; 2880, Cell Signaling Technology, Danvers, MA), and β-actin (A5216, Sigma‐Aldrich Co., St. Louis, MO), and incubated with anti- rabbit HRP-conjugated secondary antibody (1:10,000; NA934, GE Healthcare, Chicago, IL, USA) anti- mouse HRP (1:10,000; NA931, GE Healthcare).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining, Immunohistochemical staining, Binding Assay

Journal: Scientific Reports

Article Title: Porphyromonas gingivalis induces entero-hepatic metabolic derangements with alteration of gut microbiota in a type 2 diabetes mouse model

doi: 10.1038/s41598-021-97868-2

Figure Lengend Snippet: Proteome and metabolome profiles of the livers of db / db mice treated with Porphyromonas gingivalis ( Pg ) or CMC for 30 days, obtained using nano-LC/MS/MS, in triplicate. ( a ) Volcano plots of fold change in the levels of hydrophilic metabolites, lipids, and proteins, between Pg- and CMC-treated mice. Blue points represent significant increase or decrease (the relevant compounds are listed in Supplementary Tables and – ). ( b ) The levels of glucose metabolites (PEP, PGA, FUM, and MAL) were significantly higher ( P < 0.05), while the glycogen content was significantly lower ( P < 0.05), in Pg- treated mice, compared to that in CMC-treated mice. ( c ) Comparative proteome analysis of the levels of glycolysis/gluconeogenesis-related enzymes in Pg - and CMC-treated mice. ( d ) Comparative proteome analysis of enzyme synthesis and degradation during glycogen and G6P metabolism via the glycolytic pathway in Pg- and CMC-treated mice. PCK1, ALDOA, and TPI1 were significantly upregulated; while, Pgm2, DLAT, GPI1, LDHA, PDH, Gys2, AGL, and Gpi 1 were significantly downregulated ( c, d ), in the Pg -treated mice, compared to that in the CMC-treated mice. Panels ( b – d ) Data normalized and statistically analyzed as for Fig. a. n = 3, * P < 0.05, ** P < 0.01 and *** P < 0.001, versus control.

Article Snippet: Immunoblotting was performed using the following primary antibodies: PCK1 (1:1000; ab28455, Abcam, Toronto, Canada), G6PC (ab83690; Abcam), FOXO1 (1:000; 2880, Cell Signaling Technology, Danvers, MA), and β-actin (A5216, Sigma‐Aldrich Co., St. Louis, MO), and incubated with anti- rabbit HRP-conjugated secondary antibody (1:10,000; NA934, GE Healthcare, Chicago, IL, USA) anti- mouse HRP (1:10,000; NA931, GE Healthcare).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Scientific Reports

Article Title: Porphyromonas gingivalis induces entero-hepatic metabolic derangements with alteration of gut microbiota in a type 2 diabetes mouse model

doi: 10.1038/s41598-021-97868-2

Figure Lengend Snippet: Schematic presentation of the levels of hepatic glucose metabolites and the expression of rate-limiting enzymes of glucose metabolism in Pg - and CMC-treated diabetic mice. Fold changes in the glucose metabolites and enzymes involved in intrahepatic glucose metabolism in the Pg -treated mice, relative to that in the control. Red circles and lines: significant increases. Blue circles and lines: significant reductions. Metabolites that varied substantially and significantly between the Pg -treated mice and the control were identified using these criteria: log 2 fold change >|0.26| and P < 0.05, and P < 0.01. 2 KG, 2-Ketoglutaric acid; 6PG, 6-Phosphogluconic acid; Ace CoA, Acetyl-CoA; BPGA, 1,3-Bisphosphoglycerate; Cit, Citric acid; DHAP, Dihydroxyacetone phosphate; F1P, d -Fructose 1-phosphate; F6P, d -Fructose 6-phosphate; Frc, d -Fructose; G1P, d -Glucose 1-phosphate; G6P, d -Glucose 6-phosphate; GAP, Glyceraldehyde 3-phosphate; Glc, d -Glucose; Isocit, Isocitric acid; Lac, Lactic acid; MAL, Malic acid; Oxa, Oxaloacetic acid; PEP, Phosphoenolpyruvic acid; PGA, 2-Phospho- d -glyceric acid/3-Phospho- d -glyceric acid; Pyr, Pyruvic acid; Suc, Succinic acid; Suc CoA, Succinyl-CoA; UDP-Gal, UDP-alpha- d -galactose; UDP-Glc, Uridine 5′-diphosphate glucose; AGL, Amylo-1,6-glucosidase, 4-alpha-glucanotransferase; Aldoa, Fructose-bisphosphate aldolase A; Dlat, Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial; Pdhb, Pyruvate dehydrogenase E1 component subunit beta; Fh1, Fumarate hydratase-1; Gpi1, Glucose-6-phosphate isomerase 1; Gys2: Glycogen [starch] synthase 2; Ldha, L-lactate dehydrogenase; Pck1, Phosphoenolpyruvate carboxykinase; Pgm2, Phosphoglucomutase-2; Idh3a, Isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial; Suclg1, Succinyl-CoA ligase [GDP-forming] subunit alpha, mitochondrial; Sdhc, Succinate dehydrogenase (ubiquinone) cytochrome b560 subunit; Acly, ATP citrate (pro-S)-lyase; Tpi1, Triosephosphate isomerase 1.

Article Snippet: Immunoblotting was performed using the following primary antibodies: PCK1 (1:1000; ab28455, Abcam, Toronto, Canada), G6PC (ab83690; Abcam), FOXO1 (1:000; 2880, Cell Signaling Technology, Danvers, MA), and β-actin (A5216, Sigma‐Aldrich Co., St. Louis, MO), and incubated with anti- rabbit HRP-conjugated secondary antibody (1:10,000; NA934, GE Healthcare, Chicago, IL, USA) anti- mouse HRP (1:10,000; NA931, GE Healthcare).

Techniques: Expressing