Journal: Endocrinology

Article Title: G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance

doi: 10.1210/en.2016-1678

Figure Lengend Snippet: The GR stimulates G6PC2 promoter activity by displacing MafA. A, The G6PC2 GRE overlaps a binding site for MafA (24). B, Gel retardation supershift assays demonstrate that MafA binds the −182/−154 G6PC2 promoter region. The c-Maf antiserum cross-reacts with MafA (13). A representative gel is shown. C, Gel retardation competition experiments demonstrate that the order of MafA-binding affinity to the sequences shown in A is Maf SDM 2 > WT > Maf SDM 1. A representative gel is shown. D–I, Effect of promoter mutations on basal and Dex-stimulated G6PC2-luciferase fusion gene expression in βTC-3 cells. A reduction in MafA binding (Maf SDM 1) decreases basal expression but has variable effects on the Dex response. An increase in MafA binding (Maf SDM 2) increases basal expression and consistently reduces the Dex response. Results show the mean ± SEM of 3 experiments; *, P < .05 vs control. J, Effect of MafA overexpression on Dex-stimulated G6PC2-luciferase fusion gene expression in 832/13 cells. Overexpression of MafA inhibits G6PC2-luciferase fusion gene expression. Results show the mean ± SEM of 3 experiments; *, P < .05 vs control.

Article Snippet: The catalogue numbers are as follows: mouse G6pc2 , Mm00491176_m1; and mouse Slc37a4 , Mm00484574_m1.

Techniques: Activity Assay, Binding Assay, Electrophoretic Mobility Shift Assay, Luciferase, Gene Expression, Expressing, Control, Over Expression

Journal: Endocrinology

Article Title: G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance

doi: 10.1210/en.2016-1678

Figure Lengend Snippet: The rs2232316 G6PC2 SNP has opposite effects on basal and Dex-stimulated promoter activity. A–D, The influence of the alternate alleles of the rs573225 and rs2232316 SNPs on basal (A and C) and Dex-stimulated (B and D) G6PC2-luciferase fusion gene expression was analyzed by transient transfection in βTC-3 cells. The rs573225-G and rs2232316-A alleles both increase basal expression but only the rs2232316-A allele affects the Dex response. Results show the mean ± SEM of 3 experiments; *, P < .05 vs control. E, Effect of Foxa2 overexpression on Dex-stimulated G6PC2-luciferase fusion gene expression in 832/13 cells. Overexpression of Foxa2 inhibits G6PC2-luciferase fusion gene expression. Results show the mean data ± SD; *, P < .05 vs control.

Article Snippet: The catalogue numbers are as follows: mouse G6pc2 , Mm00491176_m1; and mouse Slc37a4 , Mm00484574_m1.

Techniques: Activity Assay, Luciferase, Gene Expression, Transfection, Expressing, Control, Over Expression

Journal: Endocrinology

Article Title: G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance

doi: 10.1210/en.2016-1678

Figure Lengend Snippet: Chronic Dex treatment stimulates pancreatic G6pc2 gene expression in 129SvEv mice. Effect of chronic Dex treatment on pancreatic G6pc2 and Slc37a4 (A–C) and hepatic G6pc1 and Slc37a4 (D–F) gene expression in 129SvEv mice. Pancreatic and hepatic RNA were isolated after a 6-hour fast from control mice and mice that had received daily injections of the indicated amount of Dex phosphate for 5 days. Results show the mean ± SEM of 3 experiments; *, P < .05 vs control.

Article Snippet: The catalogue numbers are as follows: mouse G6pc2 , Mm00491176_m1; and mouse Slc37a4 , Mm00484574_m1.

Techniques: Gene Expression, Isolation, Control

Journal: Endocrinology

Article Title: G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance

doi: 10.1210/en.2016-1678

Figure Lengend Snippet: G6pc2 modulates the effect of Dex on FBG and glucose tolerance in 129SvEv mice. A and B, Schematics proposing that the induction of G6pc2 expression by Dex will increase the difference in FBG between fasted WT and KO mice. The diagrams indicate that the actual values of the x-axis (glucose) may be shifted to the right (A) or left (B), relative to those in control mice, depending on the effects of Dex on other aspects of metabolism. In either case, FPI levels are predicted to not differ between WT and G6pc2 KO mice, as observed in control mice. C, Analysis of glucose tolerance using IPGTTs in 6-hour fasted conscious Dex-treated WT 129SvEv mice. Glucose tolerance was significantly different between nondiabetic Dex-treated (n = 12) and diabetic Dex-treated (n = 5) WT mice based on ANOVA and at the indicated time points based on post hoc analyses (*, P < .05). D, Analysis of glucose tolerance using IPGTTs in 6-hour fasted conscious Dex-treated KO 129SvEv mice. Results show the mean ± SEM. Glucose tolerance was significantly different between nondiabetic Dex-treated (n = 10) and diabetic Dex-treated (n = 5) KO mice based on ANOVA and at the indicated time points based on post hoc analyses (*, P < .05). E, Weight change after 4 days of Dex (D) injections in nondiabetic 129SvEv mice (WT, n = 12; KO, n = 10). Results show the mean ± SEM; *, P < .05 vs initial body weight. F, Glucose levels in 6-hour fasted conscious control (C) (WT, n = 14; KO, n = 12) or Dex-treated nondiabetic (D) (WT, n = 12; KO, n = 10) 129SvEv male mice. Results show the mean ± SEM; *, P < .05 vs control KO; **, P < .05 vs matching WT. G, Insulin levels in 6-hour fasted conscious control (C) (WT, n = 11; KO, n = 10) or Dex-treated nondiabetic (D) (WT, n = 7; KO, n = 6) 129SvEv male mice. Results show the mean ± SEM; *, P < .05 vs control WT or KO. H, Analysis of glucose tolerance using IPGTTs in 6-hour fasted conscious control (WT, n = 14; KO, n = 12) or Dex-treated (WT, n = 12; KO, n = 10) 129SvEv mice. Results show the mean ± SEM. Glucose tolerance was significantly different between control and nondiabetic Dex-treated WT mice and control and nondiabetic Dex-treated KO mice based on ANOVA and at the indicated time points based on post hoc analyses (*, P < .05). I, Analysis of glucose tolerance using IPGTTs in 6-hour fasted conscious control (WT, n = 14; KO, n = 12) or physically restrained (PR) (WT, n = 8; KO, n = 10) 129SvEv mice. Results show the mean ± SEM. Glucose tolerance was significantly different between control and physically restrained KO mice and between physically restrained WT and KO mice based on ANOVA and at the indicated time points based on post hoc analyses (*, P < .05).

Article Snippet: The catalogue numbers are as follows: mouse G6pc2 , Mm00491176_m1; and mouse Slc37a4 , Mm00484574_m1.

Techniques: Expressing, Control

Journal: Endocrinology

Article Title: G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance

doi: 10.1210/en.2016-1678

Figure Lengend Snippet: Chronic Dex treatment stimulates pancreatic G6pc2 gene expression in C57BL/6J mice. Effect of chronic Dex treatment on pancreatic G6pc2 and Slc37a4 (A–C) and hepatic G6pc1 and Slc37a4 (D–F) gene expression in C57BL/6J mice. Pancreatic and hepatic RNA were isolated after a 6-hour fast from control mice and mice that had received daily injections of the indicated amount of Dex phosphate for 5 days. Results show the mean ± SEM of 3 experiments; *, P < .05 vs control.

Article Snippet: The catalogue numbers are as follows: mouse G6pc2 , Mm00491176_m1; and mouse Slc37a4 , Mm00484574_m1.

Techniques: Gene Expression, Isolation, Control

Journal: Endocrinology

Article Title: G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance

doi: 10.1210/en.2016-1678

Figure Lengend Snippet: The induction of G6pc2 by Dex modulates FBG and glucose tolerance in C57BL/6J mice. A, Analysis of glucose tolerance using IPGTTs in 6-hour fasted conscious control (WT, n = 14; KO, n = 12) or Dex-treated (WT, n = 13; KO, n = 11) C57BL/6J male mice. Results show the mean ± SEM. Glucose tolerance was significantly different between control and Dex-treated mice and between Dex-treated WT and KO mice based on ANOVA; *, P < .05. B, Weight change after 5 days of Dex (D) injections in WT (n = 20) and KO (n = 20) C57BL/6J mice. Results show the mean ± SEM; *, P < .05 vs initial body weight. C, Glucose levels in 6-hour fasted conscious control (C) (WT, n = 12; KO, n = 22) or Dex-treated (D) (WT, n = 36; KO, n = 46) C57BL/6J male mice. Results show the mean ± SEM; *, P < .05 vs control KO; **, P < .05 vs matching WT. D, Insulin levels in 6-hour fasted conscious control (C) (WT, n = 12; KO, n = 22) or Dex-treated (D) (WT, n = 33; KO, n = 42) C57BL/6J male mice. Results show the mean ± SEM; *, P < .05 vs control WT or KO. E, Glucose levels in 24-hour fasted conscious control (C) (WT, n = 9; KO, n = 8) or 24-hour fasted conscious Dex-treated (D) (WT, n = 6; KO, n = 8) C57BL/6J male mice. Results show the mean ± SEM; *, P < .05 vs control WT or KO; **, P < .05 vs matching WT. F, Insulin levels in 24-hour fasted conscious control (C) (WT, n = 9; KO, n = 9) or 24-hour fasted conscious Dex-treated (D) (WT, n = 15; KO, n = 12) C57BL/6J male mice. Results show the mean ± SEM; *, P < .05 vs control WT or KO. G, Analysis of glucose tolerance using IPGTTs in 24-hour fasted conscious control (WT, n = 9; KO, n = 8) or Dex-treated (WT, n = 6; KO, n = 8) C57BL/6J male mice. Results show the mean ± SEM. Glucose tolerance was significantly different between control and Dex-treated KO mice based on ANOVA and at the indicated time points based on post hoc analyses (*, P < .05).

Article Snippet: The catalogue numbers are as follows: mouse G6pc2 , Mm00491176_m1; and mouse Slc37a4 , Mm00484574_m1.

Techniques: Control

Journal: PLoS Genetics

Article Title: Identification and Functional Characterization of G6PC2 Coding Variants Influencing Glycemic Traits Define an Effector Transcript at the G6PC2-ABCB11 Locus

doi: 10.1371/journal.pgen.1004876

Figure Lengend Snippet: Coding variants associated with FG and FI levels at exome-wide significance.

Article Snippet: Human G6PC2 cDNA (NM_021176.2) within a pCMV6-Entry vector (with a C-terminal FLAG-tag) was purchased from OriGene (RC211146).

Techniques: Variant Assay

Journal: PLoS Genetics

Article Title: Identification and Functional Characterization of G6PC2 Coding Variants Influencing Glycemic Traits Define an Effector Transcript at the G6PC2-ABCB11 Locus

doi: 10.1371/journal.pgen.1004876

Figure Lengend Snippet: G6PC2 gene-based association with FG levels using SKAT and BURDEN test

Article Snippet: Human G6PC2 cDNA (NM_021176.2) within a pCMV6-Entry vector (with a C-terminal FLAG-tag) was purchased from OriGene (RC211146).

Techniques:

Journal: PLoS Genetics

Article Title: Identification and Functional Characterization of G6PC2 Coding Variants Influencing Glycemic Traits Define an Effector Transcript at the G6PC2-ABCB11 Locus

doi: 10.1371/journal.pgen.1004876

Figure Lengend Snippet: rs138726309 (p.His177Tyr), rs2232323 (p.Tyr207Ser), and rs492594 (p.Val219Leu), obtained from 4,442 unrelated individuals from the Oxford Biobank. (A) Percentage minor allele frequency (MAF) and effect size estimates ( β ^ ) of the four variants reported for the minor allele in mmol/L of FG after adjustment for age, sex, and BMI. (B) Haplotypes of the four associated variants in G6PC2 revealed that the glucose-lowering Leu219 allele was carried exclusively in cis with the glucose-raising allele at the GWAS SNP. Wild-type, glucose-raising alleles are circled in blue and the mutant, glucose-lowering alleles are circled in red. Diameter of the circle is proportional to the effect size estimates. Haplotype association was performed with FG derived residuals (after adjustment for age, sex, and BMI) using the most frequent haplotype as baseline.

Article Snippet: Human G6PC2 cDNA (NM_021176.2) within a pCMV6-Entry vector (with a C-terminal FLAG-tag) was purchased from OriGene (RC211146).

Techniques: Mutagenesis, Derivative Assay

Journal: PLoS Genetics

Article Title: Identification and Functional Characterization of G6PC2 Coding Variants Influencing Glycemic Traits Define an Effector Transcript at the G6PC2-ABCB11 Locus

doi: 10.1371/journal.pgen.1004876

Figure Lengend Snippet: (A) Expression levels in HEK293 and (B) INS-1E cells were determined by western blot and densitometry analysis. The multiple bands on the western blot are likely to represent glycosylated G6PC2 protein products. Data are presented as mean ± standard error of the mean for at least three independent experiments. Significant differences are indicated as ** P <0.01; *** P <0.001; **** P <0.0001. EV, empty vector; WT, wild type. (C) Expression levels in HEK293 and INS-1E cells in the presence of proteasomal inhibitor MG-132 or lysosomal inhibitor chloroquine were determined by western blot. (D) Cellular localization in HEK293 cells was assessed by immunofluorescence microscopy. Cells were double immunostained for FLAG tag (green) and calnexin (red), and merged images with a DNA stain (blue) are shown. Images were taken with laser settings that were optimized separately for each sample. Scale bar, 10µm.

Article Snippet: Human G6PC2 cDNA (NM_021176.2) within a pCMV6-Entry vector (with a C-terminal FLAG-tag) was purchased from OriGene (RC211146).

Techniques: Expressing, Western Blot, Plasmid Preparation, Immunofluorescence, Microscopy, FLAG-tag, Staining

Journal: The Journal of endocrinology

Article Title: Potential Positive and Negative Consequences of ZnT8 Inhibition

doi: 10.1530/JOE-20-0138

Figure Lengend Snippet: Comparison of pancreatic Slc30a8 (Panels A and F), Ins1 (Panels B and G), Ins2 (Panels C and H), Slc2a2 (Panels D and I), G6pc2 (Panels E and J) and hepatic G6pc1 (Panels K and P), Slc37a4 (Panels L and Q), Gys2 (Panels M and R), Ide (Panels N and S) and Insr (Panels O and T) gene expression in 6 hour fasted 4 week and 10 week old male mice, respectively. Gene expression was quantitated relative to Ppia (cyclophilin A) expression in the indicated tissue. Results show the mean data ± S.E.M. with the genotype and number of animals indicated. *, differences between WT and KO; p < 0.05, Student’s t-test.

Article Snippet: The construction of a wild type human G6PC2 - and mouse G6pc2 -luciferase fusion genes have been previously described ( Boustead, et al. 2004 ; Martin, et al. 2002 ).

Techniques: Comparison, Gene Expression, Expressing

Journal: The Journal of endocrinology

Article Title: Potential Positive and Negative Consequences of ZnT8 Inhibition

doi: 10.1530/JOE-20-0138

Figure Lengend Snippet: Regulation of endogenous Fos (Panels A and H), Mt1a (Panels B and I), Ins2 (Panels C and J), Slc30a8 (Panels D and K) and G6pc2 (Panels E and L) gene expression over time by 250 μM zinc chloride and 20 mM calcium chloride, respectively. Gene expression was quantitated relative to Ppia (cyclophilin A) expression. Results represent the mean data ± S.E.M. derived from 3–4 independent experiments. *, differences with t=0; p < 0.05, one-way ANOVA. Regulation of mouse G6pc2-luciferase (Panel F) and metal response element-albumin-luciferase (Panel G) fusion gene expression by the indicated concentration of zinc chloride. Regulation of mouse G6pc2-luciferase (Panel M) and cAMP response element-albumin-luciferase (Panel N) fusion gene expression by the indicated concentration of calcium chloride. Results represent the mean data ± S.E.M. derived from 3 independent experiments. *, differences with no zinc or calcium chloride; p < 0.05, one-way ANOVA.

Article Snippet: The construction of a wild type human G6PC2 - and mouse G6pc2 -luciferase fusion genes have been previously described ( Boustead, et al. 2004 ; Martin, et al. 2002 ).

Techniques: Gene Expression, Expressing, Derivative Assay, Luciferase, Concentration Assay

Journal: Frontiers in Endocrinology

Article Title: A molecular signature for the G6PC3/SLC37A2/SLC37A4 interactors in glioblastoma disease progression and in the acquisition of a brain cancer stem cell phenotype

doi: 10.3389/fendo.2023.1265698

Figure Lengend Snippet: Increased gene expression of G6PC3 , SLC37A2 , and SLC37A4 in clinically-annotated glioblastoma tumor tissues. In silico analysis of transcript levels was performed for (A) three members of the G6PC family (G6PC1, G6PC2, G6PC3) and (B) four members of the SLC37A family (SLC37A1, SLC37A2, SLC37A3, SLC37A4) using RNA extracted from clinical samples from glioblastoma (GBM) and low-grade glioma (LGG) (red boxes) and compared to healthy tissue (grey boxes). (* p < 0.05).

Article Snippet: The following QuantiTect primer sets were provided by QIAGEN: G6PC1 (Hs_G6PC_1_SG QT00031913), G6PC2 (Hs_G6PC2_va.1_SG QT01664152), G6PC3 (Hs_G6PC3_1_SG QT00033453), SLC37A1 (SLC37A1_1_SG QT00073094), SLC37A2 (Hs_SLC37A2_1_SG QT00056203), SLC37A3 (Hs_SLC37A3_1_SG QT00057148), SLC37A4 (Hs_SLC37A4_1_SG QT00024325), GAPDH (Hs_GAPDH_2_SG QT01192646), β-actin (Hs_Actb_2_SG QT01680476) and PPIA (Hs_PPIA_4_SG QT01866137).

Techniques: Expressing, In Silico