Journal: Frontiers in Nutrition

Article Title: High-fat diet mouse model receiving L-glucose supplementations propagates liver injury

doi: 10.3389/fnut.2024.1469952

Figure Lengend Snippet: Metabolic profile assessment: (A) Fasting blood sugar (FBS) was assessed as indicated in materials and methods. GLUT2 expressions were quantitated by (B) western blot and (C) RT-PCR methods. (D) Serum insulin C-peptide measured by ELISA. (E) Hepatic expression of inulin receptor (IR) was evaluated from liver sections by western blot (F) HOMA2 calculator calculated HOMA-IR. Paired and unpaired Student’s t -test and ANOVA were used for statistically significant differences. p value was compared between RD and HFD control groups or within RD or HFD groups. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Rabbit anti-Human/Mouse GYS2 (Proteintech, 22371-1-AP), Rabbit anti-Human/Mouse Glycogen synthase [p Ser641] (Novus bio, NBP2-67315), rabbit anti-human/mouse PYGL antibody (Proteintech, 15851-1-AP), rabbit anti-human/mouse Glut2 polyclonal antibody (Proteintech, 20436-1-AP), rabbit anti-human/mouse ADRP/Perilipin 2 Polyclonal antibody (Proteintech, 15294-1-AP), rabbit anti-human/mouse Alpha Smooth Muscle antibody (Novus, NBP1-30894), mice anti-human/mouse AKT antibody (R&D, MAB 2055), mice anti-human/mouse phospho-AKT antibody (R&D, MAB 887), rabbit anti-human/mouse Insulin Receptor-beta antibody (Proteintech, 20433-1-AP), and rabbit anti-human/mouse beta Actin polyclonal antibody (Proteintech, 20536-1-AP).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Maternal Heat Stress Alters Expression of Genes Associated with Nutrient Transport Activity and Metabolism in Female Placentae from Mid-Gestating Pigs

doi: 10.3390/ijms22084147

Figure Lengend Snippet: A subset of differentially expressed genes in heat stress placentae ( n = 5) compared to control placentae ( n = 5).

Article Snippet: The membranes were then incubated with anti- SLC2A2 rabbit (at 1 μg/mL, Cat# ARP41706_P050, AVIVA Systems Biology, San Diego, CA, USA) and anti- SLC15A1 mouse (1:500 dilution, Cat# sc-373742, Santa Cruz Biotechnology, Dallas, TX, USA) primary antibodies diluted in TBST containing 5% non-fat dry milk overnight at 4 °C.

Techniques: Control, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Maternal Heat Stress Alters Expression of Genes Associated with Nutrient Transport Activity and Metabolism in Female Placentae from Mid-Gestating Pigs

doi: 10.3390/ijms22084147

Figure Lengend Snippet: Relative protein expression of glucose transporter 2 (GLUT2; ( A )) and peptide transporter 1 (PEPT1; ( B )) in placentae between control (CON, n = 5) and heat stress (HS, n = 5) groups. Target protein expression was normalised to total protein loaded and expressed as normalised signals (arbitrary units). * p < 0.05. Error bars: standard error of the means.

Article Snippet: The membranes were then incubated with anti- SLC2A2 rabbit (at 1 μg/mL, Cat# ARP41706_P050, AVIVA Systems Biology, San Diego, CA, USA) and anti- SLC15A1 mouse (1:500 dilution, Cat# sc-373742, Santa Cruz Biotechnology, Dallas, TX, USA) primary antibodies diluted in TBST containing 5% non-fat dry milk overnight at 4 °C.

Techniques: Expressing, Control

Journal: Biomolecules

Article Title: Normal β-Cell Glut2 Expression Is not Required for Regulating Glucose-Stimulated Insulin Secretion and Systemic Glucose Homeostasis in Mice

doi: 10.3390/biom13030540

Figure Lengend Snippet: Generation and validation of β-cell Glut2 knockdown (β- Glut2 KD) mice. Representative genotypes of mice used in this study ( A ). Het, heterozygous; Hom, homozygous for Glut2 loxP . Std, standard DNA ladder; NC, negative control; bp, base pair. Results from qRT-PCR showing knockdown of islet Glut2 in 8 weeks old male β- Glut2 KD mice one week after inducing β-cell Glut2 deficiency, n = 6 or 7 ( B ). Immunofluorescence demonstrating deficiency of β-cell GLUT2 in male β- Glut2 KD mice, n = 6 or 7 ( C ). Three islets per section and 3 sections per mouse were quantified ( C ). Scale bar is 50 µm. Ctrl, control Glut2 loxP/loxP mice; AU, arbitrary unit. Student’s unpaired t -test was used for comparisons, *** p < 0.001. Error bars are mean ± SEM.

Article Snippet: The tissue sections were then processed for GLUT1, GLUT2, or insulin staining using previously validated antibodies [ , ]: rabbit anti-GLUT1 (1:250 dilution in TBST, ab652 or ab115730, Abcam, Waltham, MA, USA), rabbit anti-GLUT2 (1:250 dilution in TBST, 600-401-GN3, Rockland Immunochemicals, Pottstown, PA, USA), or mouse anti-insulin (1:100 dilution in TBST, sc-8033, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) incubated at 4 °C overnight.

Techniques: Negative Control, Quantitative RT-PCR, Immunofluorescence

Journal: Biomolecules

Article Title: Normal β-Cell Glut2 Expression Is not Required for Regulating Glucose-Stimulated Insulin Secretion and Systemic Glucose Homeostasis in Mice

doi: 10.3390/biom13030540

Figure Lengend Snippet: β-cell Glut2 knockdown (β- Glut2 KD) mice have normal glucose-stimulated insulin secretion. Fasting (6 h) plasma insulin levels (0 min) and glucose-stimulated insulin secretion (20 min) in male, n = 6 or 7 ( A ) and female, n = 8, 10, or 11 ( B ) mice 4 and 8 weeks after inducing β-cell Glut2 deficiency. The mice were 8–10 weeks old when β-cell Glut2 deficiency was induced. Repeated measures two-way ANOVA followed by Bonferroni’s multiple comparison test were used for comparisons. ** p < 0.01, *** p < 0.001 vs. their corresponding baseline (0 min) groups. Error bars are mean ± SEM.

Article Snippet: The tissue sections were then processed for GLUT1, GLUT2, or insulin staining using previously validated antibodies [ , ]: rabbit anti-GLUT1 (1:250 dilution in TBST, ab652 or ab115730, Abcam, Waltham, MA, USA), rabbit anti-GLUT2 (1:250 dilution in TBST, 600-401-GN3, Rockland Immunochemicals, Pottstown, PA, USA), or mouse anti-insulin (1:100 dilution in TBST, sc-8033, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) incubated at 4 °C overnight.

Techniques:

Journal: Biomolecules

Article Title: Normal β-Cell Glut2 Expression Is not Required for Regulating Glucose-Stimulated Insulin Secretion and Systemic Glucose Homeostasis in Mice

doi: 10.3390/biom13030540

Figure Lengend Snippet: β-cell Glut2 knockdown (β- Glut2 KD) mice exhibit normal glucose tolerance. Results obtained from oral glucose tolerance tests in male, n = 9 or 12 ( A , B ) and female, n = 6–10 ( C , D ) mice 4 ( A , C ) and 8 ( B , D ) weeks after inducing β-cell Glut2 deficiency. Area under the curve (AUC) is shown in inset bar graphs. The mice were 8–10 weeks old when β-cell Glut2 deficiency was induced. Immunofluorescence, n = 6 ( E ) and qRT-PCR, n = 6 ( F ) demonstrating upregulation of islet Glut1 in female β- Glut2 KD mice after completion of the study. 3 islets per section and 3 sections per mouse were quantified ( E ). Ins +ve and −ve, quantification of GLUT1 fluorescence in insulin positive and negative staining. Scale bar is 50 µm. Control, Glut2 loxP/loxP mice; AU, arbitrary unit. Student’s unpaired t -test was used for comparisons, * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars are mean ± SEM.

Article Snippet: The tissue sections were then processed for GLUT1, GLUT2, or insulin staining using previously validated antibodies [ , ]: rabbit anti-GLUT1 (1:250 dilution in TBST, ab652 or ab115730, Abcam, Waltham, MA, USA), rabbit anti-GLUT2 (1:250 dilution in TBST, 600-401-GN3, Rockland Immunochemicals, Pottstown, PA, USA), or mouse anti-insulin (1:100 dilution in TBST, sc-8033, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) incubated at 4 °C overnight.

Techniques: Immunofluorescence, Quantitative RT-PCR, Fluorescence, Negative Staining