Journal: Biomolecules

Article Title: Gintonin Stimulates Glucose Uptake in Myocytes: Involvement of Calcium and Extracellular Signal-Regulated Kinase Signaling.

doi: 10.3390/biom14101316

Figure Lengend Snippet: Figure 5. Effect of gintonin on GLUT4 expression in total lysates and plasma membrane fractions of C2C12 myotubes. (A) GLUT4 expression in total lysates. (B) GLUT4 expression in the plasma membrane fraction. C2C12 myotubes were treated with gintonin (GT, 10 µg/mL) for 120 min or insulin (INS, 100 nM) for 30 min. GLUT4 expression in total lysates and plasma membrane fraction of C2C12 myotubes was detected by immunoblotting. β-actin and Na+/K+ ATPase were also detected as loading controls. All data are shown as the mean ± SEM (n = 4); ** p < 0.01; *** p < 0.001 vs. untreated control cells (Con). Original western blot images can be found in Supplementary File S1.

Article Snippet: The cell membrane fraction and total cell lysate were also used to measure GLUT4 expression by immunoblotting using a mouse anti-GLUT4 monoclonal antibody (Santa Cruz Biotechnology) and a goat anti-mouse IgG antibody conjugated to HRP (Santa Cruz Biotechnology).

Techniques: Expressing, Clinical Proteomics, Membrane, Western Blot, Control

Journal: Biomolecules

Article Title: Gintonin Stimulates Glucose Uptake in Myocytes: Involvement of Calcium and Extracellular Signal-Regulated Kinase Signaling.

doi: 10.3390/biom14101316

Figure Lengend Snippet: Figure 6. Possible signaling pathways of gintonin (GT)-induced glucose uptake in C2C12 myotubes. Gintonin induces transient increases in intracellular calcium concentrations and ERK activation via LPA receptor (LPAR) activation. These may lead to increases in the expression and translocation of GLUT4, subsequently increasing glucose uptake. PLC, phospholipase C; ERK, extracellular signal-regulated kinase; GLUT4, glucose transporter type 4.

Article Snippet: The cell membrane fraction and total cell lysate were also used to measure GLUT4 expression by immunoblotting using a mouse anti-GLUT4 monoclonal antibody (Santa Cruz Biotechnology) and a goat anti-mouse IgG antibody conjugated to HRP (Santa Cruz Biotechnology).

Techniques: Protein-Protein interactions, Activation Assay, Expressing, Translocation Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Genetic deletion of zinc transporter ZnT 3 induces progressive cognitive deficits in mice by impairing dendritic spine plasticity and glucose metabolism

doi: 10.3389/fnmol.2024.1375925

Figure Lengend Snippet: ZnT 3 deletion is associated with a significant impairment of insulin signaling in the hippocampus. (A) Body mass analysis of ZnT 3 - / - and WT mice. *, ### Denote significant difference between groups indicated by the connector lines, p < 0.05, p < 0.001, two-way ANOVA with LSD post-hoc test. (B) Brain weight analysis of ZnT 3 - / - and WT mice. Two-way ANOVA with LSD post-hoc test. (C, D) The blood glucose levels in male (C) and female (D) mice (ZnT 3 - / - ; and WT) after fasting; n = 6 for both ZnT 3 - / - and n = 5 for both WT groups. Two-way ANOVA with LSD post-hoc test. (E, F) mRNA expression levels of Slc2a1 (GLUT1), Slc2a3 (GLUT3), and Slc2a4 (GLUT4) in cortex (E) ; and Slc2a1 (GLUT1), Slc2a3 (GLUT3), and Slc2a4 (GLUT4) and Insr (INSR) in hippocampus (F) . Expression levels are normalized to housekeeping gene Gapdh . * Denotes significant difference between groups indicated by the connector lines, p < 0.05, Mann-Whitney test. (G, H) Immunofluorescence staining of hippocampal sections of 9-months old mice. (G) : GLUT3-(red), DAPI-(blue) and GFAP (green). (H) : GLUT4-(green), DAPI-(blue) and NeuN (red). Scale bar, 50 μm; Zoom Scale bar, 5 μm. (I, J) Statistical analysis of images as those shown in (G) and (H) . * Denotes significant difference between groups indicated by the connector lines, p < 0.05, Mann-Whitney test.

Article Snippet: The following primary antibodies were used: rabbit anti-GLUT3 ployclonal antibody (20403-1-AP, Proteintech, USA; 1:2,000); mouse anti-GLUT4 monoclonal antibody (66846-1-lg, Proteintech, USA; 1:2,000); rabbit anti-GSK-3β monoclonal antibody (D5C5Z, Cell Signaling, USA; 1:1,000); and anti-GAPDH recombinant rabbit monoclonal antibody (SA30-01, HUABIO, China; 1:5,000).

Techniques: Expressing, MANN-WHITNEY, Immunofluorescence, Staining

Journal: Juntendo Medical Journal

Article Title: Maternal Protein Restriction Inhibits Insulin Signaling and Insulin Resistance in the Skeletal Muscle of Young Adult Rats

doi: 10.14789/jmj.JMJ23-0029-OA

Figure Lengend Snippet: Results of real-time polymerase chain reaction analysis of protein kinase B ( Akt2 ), phosphoinositide 3-kinases ( PIK3 ), insulin receptor substrate 1 ( IRS1 ), and glucose transporter type 4 ( GLUT4 ) mRNA in the (A) soleus muscle and (B) plantar muscle of fetal growth restriction (FGR) and control rats. ※ p < 0.05 vs. control rats.

Article Snippet: Polyvinylidene fluoride (PVDF) membrane was blocked with Bullet Blocking One for western blotting (cat. no. 13779-56; Nacalai Tesque) for 5 min, and then incubated overnight at 5°C with the following primary antibodies: rabbit anti-Akt2 monoclonal antibody (1:1000; cat. no. 9272s; Cell Signaling Technology, Danvers, MA, USA), mouse anti GLUT4 monoclonal antibody (1:1000; cat. no. 2213; Cell Signaling Technology), and rabbit anti-GAPDH monoclonal antibody (1:10000; cat. no. 5174S; Cell Signaling Technology).

Techniques: Real-time Polymerase Chain Reaction, Control

Journal: Juntendo Medical Journal

Article Title: Maternal Protein Restriction Inhibits Insulin Signaling and Insulin Resistance in the Skeletal Muscle of Young Adult Rats

doi: 10.14789/jmj.JMJ23-0029-OA

Figure Lengend Snippet: Results of western blot analysis of protein kinase B (Akt2), phosphoinositide 3-kinases (PIK3), and glucose transporter type 4 (GLUT4) in the (A) soleus muscle and (B) plantar muscle of fetal growth restriction (FGR) rats and control rats. ※ p < 0.05 vs. control rats.

Article Snippet: Polyvinylidene fluoride (PVDF) membrane was blocked with Bullet Blocking One for western blotting (cat. no. 13779-56; Nacalai Tesque) for 5 min, and then incubated overnight at 5°C with the following primary antibodies: rabbit anti-Akt2 monoclonal antibody (1:1000; cat. no. 9272s; Cell Signaling Technology, Danvers, MA, USA), mouse anti GLUT4 monoclonal antibody (1:1000; cat. no. 2213; Cell Signaling Technology), and rabbit anti-GAPDH monoclonal antibody (1:10000; cat. no. 5174S; Cell Signaling Technology).

Techniques: Western Blot, Control

Journal: Juntendo Medical Journal

Article Title: Maternal Protein Restriction Inhibits Insulin Signaling and Insulin Resistance in the Skeletal Muscle of Young Adult Rats

doi: 10.14789/jmj.JMJ23-0029-OA

Figure Lengend Snippet: Association between Akt2 and GLUT4 expression and the incremental area under the curve (iAUC) in the plantar muscle at 12 weeks of age.

Article Snippet: Polyvinylidene fluoride (PVDF) membrane was blocked with Bullet Blocking One for western blotting (cat. no. 13779-56; Nacalai Tesque) for 5 min, and then incubated overnight at 5°C with the following primary antibodies: rabbit anti-Akt2 monoclonal antibody (1:1000; cat. no. 9272s; Cell Signaling Technology, Danvers, MA, USA), mouse anti GLUT4 monoclonal antibody (1:1000; cat. no. 2213; Cell Signaling Technology), and rabbit anti-GAPDH monoclonal antibody (1:10000; cat. no. 5174S; Cell Signaling Technology).

Techniques: Expressing

Journal: Juntendo Medical Journal

Article Title: Maternal Protein Restriction Inhibits Insulin Signaling and Insulin Resistance in the Skeletal Muscle of Young Adult Rats

doi: 10.14789/jmj.JMJ23-0029-OA

Figure Lengend Snippet: DNA methylation profiles in the promoter region of CpG sites of GLUT4 in fetal growth restriction (FGR) and control rats (A). The methylation rate of GLUT4 was not significantly different between the groups (B).

Article Snippet: Polyvinylidene fluoride (PVDF) membrane was blocked with Bullet Blocking One for western blotting (cat. no. 13779-56; Nacalai Tesque) for 5 min, and then incubated overnight at 5°C with the following primary antibodies: rabbit anti-Akt2 monoclonal antibody (1:1000; cat. no. 9272s; Cell Signaling Technology, Danvers, MA, USA), mouse anti GLUT4 monoclonal antibody (1:1000; cat. no. 2213; Cell Signaling Technology), and rabbit anti-GAPDH monoclonal antibody (1:10000; cat. no. 5174S; Cell Signaling Technology).

Techniques: DNA Methylation Assay, Control, Methylation