Journal: Experimental and Therapeutic Medicine

Article Title: Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway in vitro

doi: 10.3892/etm.2025.12956

Figure Lengend Snippet: Level of SLC2A4 in HG-induced HTR-8/SVneo cells. (A) A screen of targeted genes related to gestational diabetes mellitus in the CTD, DisGeNet and Genecards databases. (B) The level of LDH was analyzed using an LDH assay. (C) The expression of SLC2A4 was analyzed by reverse transcription-quantitative PCR. (D) The expression of SLC2A4 was analyzed by western blotting. ** P<0.01 and *** P<0.001 vs. control. CTD, Comparative Toxicogenomics Database; SLC2A4, solute carrier family 2 member 4; LDH, lactate dehydrogenase; HG, high glucose.

Article Snippet: The membranes were then incubated overnight at 4 ̊C with primary antibodies against SLC2A4 (cat. no. 2213S; Cell Signaling Technology, Inc.), INSR (cat. no. 3025S; Cell Signaling Technology, Inc.), INS (cat. no. ab181547; Abcam), phosphorylated (p)-FoxO1 (cat. no. 9461S; Cell Signaling Technology, Inc.), FoxO1 (cat. no. 2880S; Cell Signaling Technology, Inc.), p-FoxO3a (cat. no. 13129S; Cell Signaling Technology, Inc.), FoxO3a (cat. no. 12829S; Cell Signaling Technology, Inc.) and GAPDH (cat. no. 97166; Cell Signaling Technology, Inc.) (all 1:1,000 dilution).

Techniques: Lactate Dehydrogenase Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway in vitro

doi: 10.3892/etm.2025.12956

Figure Lengend Snippet: Effect of SLC2A4 overexpression on HG-induced HTR-8/SVneo cells. The expression of SLC2A4 was analyzed by (A) RT-qPCR and (B) western blotting. The expression of INS and INSR was analyzed by (C) RT-qPCR and (D) western blotting. (E) Glucose uptake was detected using a glucose uptake assay. (F) Cell viability was measured using a Cell Counting Kit-8 assay. *** P<0.001 vs. Control; ns P>0.05 vs. HG; # P<0.05 and ### P<0.001 vs. HG + pc-NC. SLC2A4, solute carrier family 2 member 4; HG, high glucose; RT-qPCR, reverse transcription-quantitative PCR; ns, not significant; NC, negative control; OD, optical density; INS, insulin; INSR, INS receptor.

Article Snippet: The membranes were then incubated overnight at 4 ̊C with primary antibodies against SLC2A4 (cat. no. 2213S; Cell Signaling Technology, Inc.), INSR (cat. no. 3025S; Cell Signaling Technology, Inc.), INS (cat. no. ab181547; Abcam), phosphorylated (p)-FoxO1 (cat. no. 9461S; Cell Signaling Technology, Inc.), FoxO1 (cat. no. 2880S; Cell Signaling Technology, Inc.), p-FoxO3a (cat. no. 13129S; Cell Signaling Technology, Inc.), FoxO3a (cat. no. 12829S; Cell Signaling Technology, Inc.) and GAPDH (cat. no. 97166; Cell Signaling Technology, Inc.) (all 1:1,000 dilution).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Cell Counting, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Enhanced SLC2A4 expression mitigates insulin resistance in gestational diabetes mellitus via the FoxO signaling pathway in vitro

doi: 10.3892/etm.2025.12956

Figure Lengend Snippet: Role of the FoxO signaling pathway in pc-SLC2A4-induced changes in HG-induced HTR-8/SVneo cells. (A) Screen of targeted pathways related to gestational diabetes mellitus and SLC2A4 (five common genes identified through the CTD/GeneCards/DisGeNet database) using Kyoto Encyclopedia of Genes and Genomes, suggesting a critical role for FoxO signaling pathway. (B) The expression levels of p-FoxO1, FoxO1, p-FoxO3a and FoxO3a were analyzed by western blotting, GAPDH was used as the internal control for the results. The expression of INS and INSR was analyzed by (C) reverse transcription-quantitative PCR and (D) western blotting. (E) Glucose uptake was detected by glucose uptake assay. (F) Cell viability was measured using a Cell Counting Kit-8 assay. *** P<0.001 vs. Control; ns P>0.05 vs. HG; ### P<0.001 vs. HG + pc-NC; $ P<0.05, $$ P<0.01 and $$$ P<0.001 vs. HG + pc-SLC2A4. SLC2A4, solute carrier family 2 member 4; HG, high glucose; ns, not significant; NC, negative control; OD, optical density; INS, insulin; INSR, INS receptor; p, phosphorylated; FoxO, Forkhead box O.

Article Snippet: The membranes were then incubated overnight at 4 ̊C with primary antibodies against SLC2A4 (cat. no. 2213S; Cell Signaling Technology, Inc.), INSR (cat. no. 3025S; Cell Signaling Technology, Inc.), INS (cat. no. ab181547; Abcam), phosphorylated (p)-FoxO1 (cat. no. 9461S; Cell Signaling Technology, Inc.), FoxO1 (cat. no. 2880S; Cell Signaling Technology, Inc.), p-FoxO3a (cat. no. 13129S; Cell Signaling Technology, Inc.), FoxO3a (cat. no. 12829S; Cell Signaling Technology, Inc.) and GAPDH (cat. no. 97166; Cell Signaling Technology, Inc.) (all 1:1,000 dilution).

Techniques: Expressing, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: α-Pinene, a Main Component of Pinus Essential Oils, Enhances the Expression of Insulin-Sensitive Glucose Transporter Type 4 in Murine Skeletal Muscle Cells

doi: 10.3390/ijms25021252

Figure Lengend Snippet: Effects of the Pinus spp. essential oils on Glut4 mRNA expression. ( A ) C2C12 cells were exposed to increasing doses of the P. radiata (PrEO) or P. nigra (PnEO) essential oils for 72 h; cell viability was determined by MTT assay. ( B ) RT-qPCR analysis of Glut4 mRNA expression in C2C12 cells treated with the PrEO and PnEO. ( C ) The PnEO regulated the mRNA expression of Glut4 in C2C12 cells in a dose-dependent manner. Asterisks indicate significant values with respect to control untreated cells ( p < 0.05).

Article Snippet: Whole cells and cells permeabilized by cold absolute methanol for 15 min were stained with antibodies against GLUT4 (Origene, Rockville, MD, USA) (1:100) and then with phycoerythrin-conjugated anti-rabbit secondary antibodies (1:200) (Thermofisher, Milan, Italy).

Techniques: Expressing, MTT Assay, Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Pinene, a Main Component of Pinus Essential Oils, Enhances the Expression of Insulin-Sensitive Glucose Transporter Type 4 in Murine Skeletal Muscle Cells

doi: 10.3390/ijms25021252

Figure Lengend Snippet: Treatment with the PnEO increases the number of GLUT4 transporters present on the C2C12 cell surface. Data are reported as median fluorescence values of the populations. ( A ) Dose-dependent effect of human insulin on the exposure of GLUT4 on the cell surface. ( B ) Dose-dependent effect of the PnEO on the exposure of GLUT4 on the cell surface. INS100, 100 nM insulin. ( C ) Dose-dependent effect of the PnEO on total GLUT4 expression (cell surface + GSV). INS100, 100 nM insulin. Asterisks indicate significant values relative to control untreated cells ( p < 0.05).

Article Snippet: Whole cells and cells permeabilized by cold absolute methanol for 15 min were stained with antibodies against GLUT4 (Origene, Rockville, MD, USA) (1:100) and then with phycoerythrin-conjugated anti-rabbit secondary antibodies (1:200) (Thermofisher, Milan, Italy).

Techniques: Fluorescence, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Pinene, a Main Component of Pinus Essential Oils, Enhances the Expression of Insulin-Sensitive Glucose Transporter Type 4 in Murine Skeletal Muscle Cells

doi: 10.3390/ijms25021252

Figure Lengend Snippet: Effects of principal terpenes found in the Pinus essential oils on GLUT4 expression. ( A ) RT-qPCR analysis of Glut4 mRNA expression in C2C12 cells treated with α-pinene, β-pinene, and eucalyptol. ( B ) A representative flow cytometric analysis of GLUT4 protein in C2C12 cells treated with α-pinene and the PnEO or in untreated cells. ( C , D ) Hexokinase I activity in C2C12 cells treated with α-pinene in ( C ) and the PnEO in ( D ). Asterisks indicate significant values relative to control untreated cells ( p < 0.05).

Article Snippet: Whole cells and cells permeabilized by cold absolute methanol for 15 min were stained with antibodies against GLUT4 (Origene, Rockville, MD, USA) (1:100) and then with phycoerythrin-conjugated anti-rabbit secondary antibodies (1:200) (Thermofisher, Milan, Italy).

Techniques: Expressing, Quantitative RT-PCR, Activity Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Pinene, a Main Component of Pinus Essential Oils, Enhances the Expression of Insulin-Sensitive Glucose Transporter Type 4 in Murine Skeletal Muscle Cells

doi: 10.3390/ijms25021252

Figure Lengend Snippet: Expression of Glut4 in differentiated C2C12 cells. ( A ) Representative light microscopy images of three experiments reproduced in triplicate of undifferentiated (day zero) and differentiated (day 14) C2C12 cells. ( B ) RT-qPCR analysis of Glut4 mRNA expression in C2C12-differentiated cells. The asterisk indicates the significant value obtained after differentiation compared to untreated control cells ( p < 0.05).

Article Snippet: Whole cells and cells permeabilized by cold absolute methanol for 15 min were stained with antibodies against GLUT4 (Origene, Rockville, MD, USA) (1:100) and then with phycoerythrin-conjugated anti-rabbit secondary antibodies (1:200) (Thermofisher, Milan, Italy).

Techniques: Expressing, Light Microscopy, Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: α-Pinene, a Main Component of Pinus Essential Oils, Enhances the Expression of Insulin-Sensitive Glucose Transporter Type 4 in Murine Skeletal Muscle Cells

doi: 10.3390/ijms25021252

Figure Lengend Snippet: RT-qPCR analysis of Glut4 mRNA expression in C2C12 cells in different experimental settings. ( A , B ) Effects of the PnEO in ( A ) and α-pinene in ( B ) addition at day zero of differentiation. ( C , D ) Effects of the PnEO ( C ) and α-pinene ( D ) addition after C2C12 myogenic differentiation. The level of transcripts, normalized to the housekeeping gene, refers to control untreated cells (grey column). Asterisks indicate significant values relative to control untreated cells ( p < 0.05).

Article Snippet: Whole cells and cells permeabilized by cold absolute methanol for 15 min were stained with antibodies against GLUT4 (Origene, Rockville, MD, USA) (1:100) and then with phycoerythrin-conjugated anti-rabbit secondary antibodies (1:200) (Thermofisher, Milan, Italy).

Techniques: Quantitative RT-PCR, Expressing, Control