g6pase  (Santa Cruz Biotechnology)

Journal: Current Biology

Article Title: Effective Glucose Uptake by Human Astrocytes Requires Its Sequestration in the Endoplasmic Reticulum by Glucose-6-Phosphatase-β

doi: 10.1016/j.cub.2018.08.060

Figure Lengend Snippet:

Article Snippet: Rabbit anti-G6Pase-β (IC) , Santa Cruz Biotechnology, Dallas, USA , Cat#sc-134714; RRID: AB_10647643.

Techniques: Immunocytochemistry, Western Blot, Bacteria, Virus, Plasmid Preparation, Recombinant, ATP Assay, shRNA, Software

Journal: Current Biology

Article Title: Effective Glucose Uptake by Human Astrocytes Requires Its Sequestration in the Endoplasmic Reticulum by Glucose-6-Phosphatase-β

doi: 10.1016/j.cub.2018.08.060

Figure Lengend Snippet:

Article Snippet: Quantitect primer assay: G6Pase-β , QIAGEN , Cat#Hs_G6PC3_1_SG.

Techniques: Immunocytochemistry, Western Blot, Bacteria, Virus, Plasmid Preparation, Recombinant, ATP Assay, shRNA, Software

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Primer sequences of target genes

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques:

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Primary antibodies used for immunofluorescence

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques:

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Immunofluorescence staining of proliferation and differentiation biomarkers in typical (A), ISC (B), enterocyte (C), and goblet (D) organoids. Nuclei are stained blue. Organoids were stained with stem cell marker OLFM4 (red), Paneth cell marker lysozyme (LYZ; green), enterocyte marker sucrase isomaltase (SI; red), goblet cell marker mucin 2 (MUC2; green), or enteroendocrine marker chromogranin-A (CHGA; green). Representative typical, ISC, enterocyte, and goblet organoids are shown at ×60 magnification. Bars = 25 μm. Occasional basolateral staining has been shown (personal observations) to be caused largely by autofluorescence of the Matrigel. Frequency of marked cells from at least 5 organoids from 3 mice (vi of each organoid type, a,b,c,dP ≤ 0.05). Immunocytochemistry of the fructose-responsive enzyme glucose-6-phosphatase (G6Pase) in enterocyte, ISC, and secretory goblet organoids is shown (E). All organoid types were analyzed on the 7th day of experiment, which for goblet, Paneth, and enterocyte organoids, is the 4th day after differentiation (see Fig. 1). The control panel is an image of an enterocyte organoid incubated without the primary antibody.

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques: Immunofluorescence, Staining, Marker, Immunocytochemistry, Control, Incubation

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Effect of cell type on the mRNA expression of classical fructose-responsive genes Glut5, G6Pase, and Khk. Typical and ISC organoids were incubated in their respective culture media for 6 days after generation from crypt precursors. Enterocyte and goblet organoids were incubated in their respective culture media for 3 days after directed differentiation from ISC precursors. Stem cells with very few Paneth cells constituted ISC organoids; enterocytes made up almost entirely enterocyte organoids, with few goblet and occasional enteroendocrine cells, whereas goblet organoids consisted primarily of goblet with some enteroendocrine cells. Media in each organoid type was supplemented with an additional 5 mM glucose, 5 mM fructose, or 10 mM fructose for 24 h. Levels of mRNA in all organoids were normalized to that in typical organoids exposed to 5 mM glucose (1.0). Only typical, enterocyte, and goblet organoids responded to supplemental fructose. Expression of the control gene Sglt1 did not vary with fructose. These results were confirmed 3 other times, using ISC, enterocyte, and goblet organoids from different mice. Analysis was by 2-way ANOVA, a,b,cP ≤ 0.05 within cell type, n = 3.

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques: Expressing, Incubation, Control

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Effect of fructose incubation of organoids enriched in Paneth cells on fructose-responsive genes Glut5, G6Pase, and Khk. Because Paneth cells also arise from secretory progenitors, we made (i to iv) then challenged Paneth cell-enriched organoids with 10 mM fructose (10 mM glucose as control, v). *P < 0.05, n = 3; for i-iii, a,b,c,dP < 0.05). Fructose induced marked increases in expression of Glut5, G6Pase, Khk, and Si genes. vi: Challenging ISC-enriched organoids with 20 mM fructose to increase the concentration gradient still had no effect on glucose-responsive genes.

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques: Incubation, Control, Expressing, Concentration Assay

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Effect of glucose concentration in the incubation medium on fructose-responsive genes. The media in typical and enterocyte organoids was changed from 17.5 to 11.5 mM glucose (control: 17.5 to 17.5 mM) at the same time that fructose was added 24 h before the termination of the experiment. We added 5 mM glucose or 5 mM or 10 mM fructose to organoids in normal 17.5 mM or low 11.5 mM glucose media (results separated by a dashed line). Although there was an expected effect of fructose on expression of Glut5 (i) and G6Pase (ii), there was no effect of glucose concentration in the media. Alpi (iii) expression was independent of media and sugar in both typical and enterocyte organoids. There was also an effect of cell type on the expression of these genes that are highly expressed in enterocytes in vivo. Expression was normalized to that in 5 mM glucose of the control 17.5 to 17.5 mM experiment (1.0). Analysis was by 3-way ANOVA (see text for details), n = 3.

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques: Concentration Assay, Incubation, Control, Expressing, In Vivo

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Effect of Glut5 and Khk deletion on fructose sensing by typical organoids. Typical organoids from wild-type (WT), Glut5−/−, and Khk−/− mice were cultured for 6 days then incubated in media supplemented with 5 mM glucose, 5 mM fructose, or 10 mM fructose for 24 h on the final day of culture. There was no effect of genotype on expression of undeleted genes. Levels of mRNA in 5 and 10 mM fructose were normalized to that in 5 mM glucose (1.0) for each genotype. Except for G6Pase expression, which did not require glucose transporter 5 (GLUT5), fructose transport and metabolism were clearly necessary for fructose sensing. a,b,cP ≤ 0.05 within genotype, n = 3.

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques: Cell Culture, Incubation, Expressing

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

doi: 10.1152/ajpgi.00416.2016

Figure Lengend Snippet: Effect of CHIR-induced dedifferentiation of enterocyte organoids on expression of genes involved in the fructose response and in marking cell types. From ISC precursors, enterocyte organoids were cultured for 3 days with C59 +VPA to full differentiation as indicated by peak expression of fructose-responsive genes. To force dedifferentiation, the Wnt inhibitor C59 was removed from the medium, and enterocyte organoids were then exposed for 36 h to 3 (dENT1) or 6 μM CHIR (dENT2) or for 6 days to 6 μM CHIR (dENT3). ENT, dENT1, dENT2, dENT3, and ISC organoids from the same mouse were then challenged with 5 mM fructose in the last 12 h before harvest. Gene expression was normalized to glucose-incubated, enterocyte organoids. Although expression of fructose-responsive genes decreased markedly with CHIR treatment, Glut5 and G6Pase expression still responded to fructose incubation in all dedifferentiated enterocyte organoids. Lgr5+ levels increased almost 100-fold with CHIR-induced dedifferentiation but never reached levels observed in ISC organoids. Lyz and Muc2 expression were also affected by dedifferentiation (n = 3 per treatment).

Article Snippet: Glucose-6-phosphatase (G6PASE) , Santa Cruz Biotechnology , sc-25840 , 1:200.

Techniques: Expressing, Cell Culture, Gene Expression, Incubation

Journal: Molecular genetics and metabolism

Article Title: Functional analysis of mutations in a severe congenital neutropenia syndrome caused by glucose-6-phosphatase-β deficiency

doi: 10.1016/j.ymgme.2014.11.012

Figure Lengend Snippet: Table 1

Article Snippet: The membranes were incubated overnight with a rabbit polyclonal antibody against amino acids 77 to 114 of human G6Pase-β or a mouse monoclonal antibody against β-actin (Santa Cruz Biotechnology, Dallas, TX), washed, then incubated with the appropriate species of horseradish peroxidase-conjugated second antibody.

Techniques: Activity Assay

Journal: Physiological Reports

Article Title: Developmental regulation of key gluconeogenic molecules in nonhuman primates

doi: 10.14814/phy2.12243

Figure Lengend Snippet: Effect of gestational age on protein content of key hepatic gluconeogenic molecules in fetal baboons. PEPCK‐M (A), PEPCK‐C (B), G6Pase‐ α (C), G6Pase‐ β (D) were measured by Western blotting. n = 5 per group. Representative blots from three animals per group are also shown. d = day and GA = gestational age. Data are means ± SE. * P < 0.01.

Article Snippet: Protein content was measured using primary antibodies against the following gluconeogenic enzymes: PEPCK‐Cytosolic (PEPCK‐C), PEPCK‐Mitochondrial (PEPCK‐M), G6Pase‐ α , G6Pase‐ β , FBPase (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Western Blot

Journal: Physiological Reports

Article Title: Developmental regulation of key gluconeogenic molecules in nonhuman primates

doi: 10.14814/phy2.12243

Figure Lengend Snippet: Effect of gestational age on gene expression of key hepatic gluconeogenic molecules in fetal baboons. PEPCK‐C (A), G6Pase‐ β (B), FBPase (C) were measured by qrt‐PCR. Dotted line indicates split graph due to the high mRNA content of 175d GA animals relative to their counterparts. n = 5 per group. d = day and GA = gestational age. Data are means ± SE. * P < 0.01.

Article Snippet: Protein content was measured using primary antibodies against the following gluconeogenic enzymes: PEPCK‐Cytosolic (PEPCK‐C), PEPCK‐Mitochondrial (PEPCK‐M), G6Pase‐ α , G6Pase‐ β , FBPase (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Gene Expression, Quantitative RT-PCR

Journal: Physiological Reports

Article Title: Developmental regulation of key gluconeogenic molecules in nonhuman primates

doi: 10.14814/phy2.12243

Figure Lengend Snippet: Postnatal changes in gene expression of key gluconeogenic molecules. PEPCK‐C (A), G6Pase‐ β (B) and FOXO1 (C) were measured by qrt‐PCR. Adults ( n = 4) and neonates ( n = 20); preterm ( n = 12), term ( n = 6), d = day and GA = gestational age. Data are means ± SE. * P < 0.05 and ** P < 0.001.

Article Snippet: Protein content was measured using primary antibodies against the following gluconeogenic enzymes: PEPCK‐Cytosolic (PEPCK‐C), PEPCK‐Mitochondrial (PEPCK‐M), G6Pase‐ α , G6Pase‐ β , FBPase (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Gene Expression, Quantitative RT-PCR

Journal: Molecular genetics and metabolism

Article Title: Functional analysis of mutations in a severe congenital neutropenia syndrome caused by glucose-6-phosphatase-β deficiency

doi: 10.1016/j.ymgme.2014.11.012

Figure Lengend Snippet: Table 1

Article Snippet: The membranes were incubated overnight with a rabbit polyclonal antibody against amino acids 77 to 114 of human G6Pase-β or a mouse monoclonal antibody against β-actin (Santa Cruz Biotechnology, Dallas, TX), washed, then incubated with the appropriate species of horseradish peroxidase-conjugated second antibody.

Techniques: Activity Assay