t2dm exendin 4 group  (MedChemExpress)

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Cortical strength is weaker in mice with type 2 diabetes mellitus (T2DM) treated with metformin compared with those treated with exendin-4. (A) Schematic chart of induction of T2DM in mice (WT, GLP1RKO) and the treatments with metformin or exendin-4. WT: wild type; GLP1RKO: glucagon-like peptide-1 receptor knockout. (B) Micro-CT images representing trabecular formation in control mice, mice with T2DM, mice with T2DM treated with exendin-4/metformin, and GLP-1R knockout mice with T2DM treated with exendin-4 within a region of interest (ROI). n=6 mice per group. Con: control; T2DM: type 2 diabetes mellitus; Met: metformin; Ex-4: Exendin-4; Micro-CT: micro-computed tomography. (C) Quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 mice in each group. (D) Micro-CT images representing cortical formation in control mice, mice with T2DM, mice with T2DM treated with exendin-4/metformin and GLP-1R knockdown mice with T2DM treated with exendin-4 within an ROI, and quantitative analysis of micro-CT images. n=6 mice in each group. (E) Effects of metformin or exendin-4 on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 mice in each group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the synergistic therapeutic effects of combined Exendin-4 and metformin therapy on osteogenic differentiation, mice were randomly divided into 5 groups (n = 6/group): T2DM group, T2DM + metformin group (300 mg/kg/day, MCE), T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + metformin(300 mg/kg/day, MCE) + Exendin-4 (20 mg/kg, MCE,13241–33-3) group, and T2DM + anti-Sema4D group (200 μg/kg, sc-390675).

Techniques: Knock-Out, Micro-CT, Control, Knockdown, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 reduces Sema4D levels to elevate cortical strength in mice with type 2 diabetes mellitus (T2DM). (A) Schedule of enzyme-linked immunosorbent assay (ELISA) and the corresponding groups. (B) ELISA to determine the levels of ALP, Runx-2, OCN, OPN, and Sema4D in control mice, mice with T2DM, mice with T2DM treated with metformin (Met)/exendin-4 (Ex-4), and GLP-1R knockdown T2DM mice treated with Ex-4. n=6 mice per group. ALP: Alkaline Phosphatase; Runx-2: Runt-related transcription factor 2; OCN: Osteocalcin; OPN: Osteopontin; Sema4D: Semaphorin 4D. (C) Schedule of micro-CT to determine changes in trabecular bone. (D) Micro-CT images representing trabecular formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within a region of interest (ROI). n=6 mice per group. (E) Quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 mice per group. (F, G) Micro-CT images representing cortical bone formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 mice per group. (H) Effects of exendin-4 or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 mice per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the synergistic therapeutic effects of combined Exendin-4 and metformin therapy on osteogenic differentiation, mice were randomly divided into 5 groups (n = 6/group): T2DM group, T2DM + metformin group (300 mg/kg/day, MCE), T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + metformin(300 mg/kg/day, MCE) + Exendin-4 (20 mg/kg, MCE,13241–33-3) group, and T2DM + anti-Sema4D group (200 μg/kg, sc-390675).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Knockdown, Micro-CT, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: The combination of metformin and exendin-4 can neutralize the effect of Sema4D. (A) Schematic chart of the sample preparation of micro-CT and 3-point bending experiments to determine the effect of the combination of metformin and exendin-4. (B) Micro-CT images representing trabecular formation in control mice and in mice treated with metformin, exendin-4, the combination of metformin and exendin-4, and anti-Sema4D within a region of interest (ROI), and quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 per group. (C) Micro-CT images representing cortical formation in control mice and in mice treated with metformin, exendin-4, the combination of metformin and exendin-4, and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 per group. (D) Effects of metformin, exendin-4, the combination of metformin and exendin-4, or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the synergistic therapeutic effects of combined Exendin-4 and metformin therapy on osteogenic differentiation, mice were randomly divided into 5 groups (n = 6/group): T2DM group, T2DM + metformin group (300 mg/kg/day, MCE), T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + metformin(300 mg/kg/day, MCE) + Exendin-4 (20 mg/kg, MCE,13241–33-3) group, and T2DM + anti-Sema4D group (200 μg/kg, sc-390675).

Techniques: Sample Prep, Micro-CT, Control, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 promotes pseudopodia numbers, leading to the stretching and wider spread of BMSCs via CRMP2. (A) Cell morphology observed using fluorescence microscopy. ImageJ was used to split color channels to observe the pseudopodium in human bone mesenchymal stem cells (hBMSCs). n=5 per group. White arrows indicate the pseudopodium in BMSCs. (B) Immunofluorescence staining to determine F-actin expression in BMSCs. n=5 per group. (C) Western blotting to determine F-actin expression in different groups. n=3 per group. (D) Proteomics analysis to determine the expression of different phosphorylated proteins. (E) Cell morphology observed using fluorescence microscopy. ImageJ was used to split color channels to observe the pseudopodium in BMSCs. n=5 per group. White arrows indicate the pseudopodium in BMSCs. (F) Immunofluorescence staining showing F-actin in different groups. n=5 per group. (G) Immunofluorescence staining indicating CRMP2 expression in BMSCs. n=5 per group. CRMP2: Collapsin Response Mediator Protein 2. (H) Western blotting to determine ALP, Runx-2, and Osterix expression in BMSCs. n=3 per group. (I) Alizarin red staining to analyze mineral deposits in BMSCs with DPYSL2 KD/OE. n=3 per group. DPYSL2 KD/OE: Dihydropyrimidinase like protein 2 knockdown/overexpression. (J) Western blotting to determine pCRMP2 expression in BMSCs with Sema4D or exendin-4. n=3 per group. (K) Immunofluorescence staining to determine pCRMP2 expression in BMSCs. n=5 per group. **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the synergistic therapeutic effects of combined Exendin-4 and metformin therapy on osteogenic differentiation, mice were randomly divided into 5 groups (n = 6/group): T2DM group, T2DM + metformin group (300 mg/kg/day, MCE), T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + metformin(300 mg/kg/day, MCE) + Exendin-4 (20 mg/kg, MCE,13241–33-3) group, and T2DM + anti-Sema4D group (200 μg/kg, sc-390675).

Techniques: Fluorescence, Microscopy, Immunofluorescence, Staining, Expressing, Western Blot, Knockdown, Over Expression, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 activates bone formation via the GLP-1R/PI3K/GSK-3β/CRMP2 signaling pathway. (A) Western blotting results of pCRMP2 and F-actin expression in BMSCs with GSK-3β knockdown or overexpression. n=3 per group. GSK-3β: Glycogen Synthase Kinase 3 beta. (B) Immunofluorescence staining of F-actin and pCRMP2 expression in BMSCs. n=5 per group. (C) Western blotting to determine pCRMP2 expression after Sema4D treatment with or without GSK-3β knockdown. n=3 per group. (D) Western blotting to determine pCRMP2 expression after exendin-4 treatment with or without GSK-3β knockdown. n=3 per group. (E) Western blotting to determine the expression of pGSK-3β, pCRMP2, and F-actin after Akt knockdown or overexpression. n=3 per group. Akt: Protein Kinase B/ Ak strain transforming. (F) Western blotting to determine the expression of pGSK-3β, pCRMP2, and F-actin after PI3K knockdown or overexpression. n=3 per group. PI3K: Phosphoinositide 3-Kinase. (G) Western blotting to determine the expression of pGSK-3β after exendin-4 treatment with or without PI3K knockdown and overexpression. n=3 per group. (H) Western blotting to determine P-PI3K expression after exendin-4 treatment with or without GSK-3β knockdown and overexpression. n=3 per group. (I) Western blotting to determine pGSK-3β expression after Sema4D treatment with or without PI3K knockdown and overexpression. n=3 per group. (J) Western blotting to determine pPI3K expression after Sema4D treatment with or without GSK–3β knockdown or overexpression. n=3 per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the synergistic therapeutic effects of combined Exendin-4 and metformin therapy on osteogenic differentiation, mice were randomly divided into 5 groups (n = 6/group): T2DM group, T2DM + metformin group (300 mg/kg/day, MCE), T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + metformin(300 mg/kg/day, MCE) + Exendin-4 (20 mg/kg, MCE,13241–33-3) group, and T2DM + anti-Sema4D group (200 μg/kg, sc-390675).

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Immunofluorescence, Staining, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Metformin promotes GLP-1R expression in hBMSCs via the miR-140-3p/STAT3/miR-3657 signaling pathway. (A) Schematic illustration of the sequences for miR-3657 and the wild-type or mutated 3′-UTR of GLP-1R mRNA, and dual Rluc/Fluc luciferase luminescence intensity of GLP-1R wild-type or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with miR-3657 mimics or miR–NC mimics. Rluc: Renilla luciferase; Fluc: Firefly luciferase; HEK293: Human Embryonic Kidney 293 Cells. (B, C) Protein expression of GLP-1R in hBMSCs after treatment with miR-3657 mimics (0, 10, 20, 40 μM) and the miR-3657 inhibitor (0, 10, 20, 40 μM). (D, E) Levels of ALP, RUNX-2, and Osterix in hBMSCs treated with exendin-4 and miR-3657 mimics (0, 10, 20, 40 μM)/miR-3657 inhibitor (0, 10, 20, 40 μM). (F) Schematic illustration of the sequences for STAT3 and the wild-type or mutated 3′-UTR of miR–3657, and dual Rluc/Fluc luciferase luminescence intensity of miR-3657 wild-type or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with STAT3 or pCDNA3.1. STAT3: signal transducer and activator of transcription 3. (G) qPCR to determine miR-3657 expression in hBMSCs treated with lenti-STAT3 and siR-STAT3 in a dose-dependent manner. n=3. Lenti-: lentivirus; siR-: siRNA; qPCR: quantitative polymerase chain reaction. (H) GLP-1R levels in hBMSCs after treatment with the STAT3 agonist and miR-3657 mimics (0, 10, 20, 40 μM) or treatment with the STAT3 inhibitor and miR-3657 inhibitor (0, 10, 20, 40 μM). (I) STAT3 levels in hBMSCs treated with metformin and the miR-140-3p inhibitor. (J) qPCR to determine miR-3657 expression in hBMSCs treated with metformin and miR-140–30 inhibitor/siR-STAT3. (K) Schematic illustration of the sequences for miR-140-3p and wild-type or mutated 3′-UTR of STAT3 mRNA, and dual Rluc/Fluc luciferase luminescence intensity of STAT3 wild-type or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with miR-140-3p mimics or miR-NC mimics. (L, M) Protein expression of STAT3 in hBMSCs after treatment with miR-140-3p mimics (0, 10, 20, 40 μM) and with the miR-140-3p inhibitor (0, 10, 20, 40 μM).

Article Snippet: To investigate the synergistic therapeutic effects of combined Exendin-4 and metformin therapy on osteogenic differentiation, mice were randomly divided into 5 groups (n = 6/group): T2DM group, T2DM + metformin group (300 mg/kg/day, MCE), T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + metformin(300 mg/kg/day, MCE) + Exendin-4 (20 mg/kg, MCE,13241–33-3) group, and T2DM + anti-Sema4D group (200 μg/kg, sc-390675).

Techniques: Expressing, Luciferase, Transfection, Real-time Polymerase Chain Reaction

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Cortical strength is weaker in mice with type 2 diabetes mellitus (T2DM) treated with metformin compared with those treated with exendin-4. (A) Schematic chart of induction of T2DM in mice (WT, GLP1RKO) and the treatments with metformin or exendin-4. WT: wild type; GLP1RKO: glucagon-like peptide-1 receptor knockout. (B) Micro-CT images representing trabecular formation in control mice, mice with T2DM, mice with T2DM treated with exendin-4/metformin, and GLP-1R knockout mice with T2DM treated with exendin-4 within a region of interest (ROI). n=6 mice per group. Con: control; T2DM: type 2 diabetes mellitus; Met: metformin; Ex-4: Exendin-4; Micro-CT: micro-computed tomography. (C) Quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 mice in each group. (D) Micro-CT images representing cortical formation in control mice, mice with T2DM, mice with T2DM treated with exendin-4/metformin and GLP-1R knockdown mice with T2DM treated with exendin-4 within an ROI, and quantitative analysis of micro-CT images. n=6 mice in each group. (E) Effects of metformin or exendin-4 on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 mice in each group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To figure out the effect of Sema4D on bone formation, mice were randomly divided into 5 groups (n = 6/group): control group, T2DM group, T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + anti-Sema4D group (200 μg/kg, sc-390675), and T2DM + anti-Sema4D + Exendin-4 group.

Techniques: Knock-Out, Micro-CT, Control, Knockdown, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 reduces Sema4D levels to elevate cortical strength in mice with type 2 diabetes mellitus (T2DM). (A) Schedule of enzyme-linked immunosorbent assay (ELISA) and the corresponding groups. (B) ELISA to determine the levels of ALP, Runx-2, OCN, OPN, and Sema4D in control mice, mice with T2DM, mice with T2DM treated with metformin (Met)/exendin-4 (Ex-4), and GLP-1R knockdown T2DM mice treated with Ex-4. n=6 mice per group. ALP: Alkaline Phosphatase; Runx-2: Runt-related transcription factor 2; OCN: Osteocalcin; OPN: Osteopontin; Sema4D: Semaphorin 4D. (C) Schedule of micro-CT to determine changes in trabecular bone. (D) Micro-CT images representing trabecular formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within a region of interest (ROI). n=6 mice per group. (E) Quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 mice per group. (F, G) Micro-CT images representing cortical bone formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 mice per group. (H) Effects of exendin-4 or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 mice per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To figure out the effect of Sema4D on bone formation, mice were randomly divided into 5 groups (n = 6/group): control group, T2DM group, T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + anti-Sema4D group (200 μg/kg, sc-390675), and T2DM + anti-Sema4D + Exendin-4 group.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Knockdown, Micro-CT, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: The combination of metformin and exendin-4 can neutralize the effect of Sema4D. (A) Schematic chart of the sample preparation of micro-CT and 3-point bending experiments to determine the effect of the combination of metformin and exendin-4. (B) Micro-CT images representing trabecular formation in control mice and in mice treated with metformin, exendin-4, the combination of metformin and exendin-4, and anti-Sema4D within a region of interest (ROI), and quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 per group. (C) Micro-CT images representing cortical formation in control mice and in mice treated with metformin, exendin-4, the combination of metformin and exendin-4, and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 per group. (D) Effects of metformin, exendin-4, the combination of metformin and exendin-4, or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To figure out the effect of Sema4D on bone formation, mice were randomly divided into 5 groups (n = 6/group): control group, T2DM group, T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + anti-Sema4D group (200 μg/kg, sc-390675), and T2DM + anti-Sema4D + Exendin-4 group.

Techniques: Sample Prep, Micro-CT, Control, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 promotes pseudopodia numbers, leading to the stretching and wider spread of BMSCs via CRMP2. (A) Cell morphology observed using fluorescence microscopy. ImageJ was used to split color channels to observe the pseudopodium in human bone mesenchymal stem cells (hBMSCs). n=5 per group. White arrows indicate the pseudopodium in BMSCs. (B) Immunofluorescence staining to determine F-actin expression in BMSCs. n=5 per group. (C) Western blotting to determine F-actin expression in different groups. n=3 per group. (D) Proteomics analysis to determine the expression of different phosphorylated proteins. (E) Cell morphology observed using fluorescence microscopy. ImageJ was used to split color channels to observe the pseudopodium in BMSCs. n=5 per group. White arrows indicate the pseudopodium in BMSCs. (F) Immunofluorescence staining showing F-actin in different groups. n=5 per group. (G) Immunofluorescence staining indicating CRMP2 expression in BMSCs. n=5 per group. CRMP2: Collapsin Response Mediator Protein 2. (H) Western blotting to determine ALP, Runx-2, and Osterix expression in BMSCs. n=3 per group. (I) Alizarin red staining to analyze mineral deposits in BMSCs with DPYSL2 KD/OE. n=3 per group. DPYSL2 KD/OE: Dihydropyrimidinase like protein 2 knockdown/overexpression. (J) Western blotting to determine pCRMP2 expression in BMSCs with Sema4D or exendin-4. n=3 per group. (K) Immunofluorescence staining to determine pCRMP2 expression in BMSCs. n=5 per group. **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To figure out the effect of Sema4D on bone formation, mice were randomly divided into 5 groups (n = 6/group): control group, T2DM group, T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + anti-Sema4D group (200 μg/kg, sc-390675), and T2DM + anti-Sema4D + Exendin-4 group.

Techniques: Fluorescence, Microscopy, Immunofluorescence, Staining, Expressing, Western Blot, Knockdown, Over Expression, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 activates bone formation via the GLP-1R/PI3K/GSK-3β/CRMP2 signaling pathway. (A) Western blotting results of pCRMP2 and F-actin expression in BMSCs with GSK-3β knockdown or overexpression. n=3 per group. GSK-3β: Glycogen Synthase Kinase 3 beta. (B) Immunofluorescence staining of F-actin and pCRMP2 expression in BMSCs. n=5 per group. (C) Western blotting to determine pCRMP2 expression after Sema4D treatment with or without GSK-3β knockdown. n=3 per group. (D) Western blotting to determine pCRMP2 expression after exendin-4 treatment with or without GSK-3β knockdown. n=3 per group. (E) Western blotting to determine the expression of pGSK-3β, pCRMP2, and F-actin after Akt knockdown or overexpression. n=3 per group. Akt: Protein Kinase B/ Ak strain transforming. (F) Western blotting to determine the expression of pGSK-3β, pCRMP2, and F-actin after PI3K knockdown or overexpression. n=3 per group. PI3K: Phosphoinositide 3-Kinase. (G) Western blotting to determine the expression of pGSK-3β after exendin-4 treatment with or without PI3K knockdown and overexpression. n=3 per group. (H) Western blotting to determine P-PI3K expression after exendin-4 treatment with or without GSK-3β knockdown and overexpression. n=3 per group. (I) Western blotting to determine pGSK-3β expression after Sema4D treatment with or without PI3K knockdown and overexpression. n=3 per group. (J) Western blotting to determine pPI3K expression after Sema4D treatment with or without GSK–3β knockdown or overexpression. n=3 per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To figure out the effect of Sema4D on bone formation, mice were randomly divided into 5 groups (n = 6/group): control group, T2DM group, T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + anti-Sema4D group (200 μg/kg, sc-390675), and T2DM + anti-Sema4D + Exendin-4 group.

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Immunofluorescence, Staining, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Metformin promotes GLP-1R expression in hBMSCs via the miR-140-3p/STAT3/miR-3657 signaling pathway. (A) Schematic illustration of the sequences for miR-3657 and the wild-type or mutated 3′-UTR of GLP-1R mRNA, and dual Rluc/Fluc luciferase luminescence intensity of GLP-1R wild-type or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with miR-3657 mimics or miR–NC mimics. Rluc: Renilla luciferase; Fluc: Firefly luciferase; HEK293: Human Embryonic Kidney 293 Cells. (B, C) Protein expression of GLP-1R in hBMSCs after treatment with miR-3657 mimics (0, 10, 20, 40 μM) and the miR-3657 inhibitor (0, 10, 20, 40 μM). (D, E) Levels of ALP, RUNX-2, and Osterix in hBMSCs treated with exendin-4 and miR-3657 mimics (0, 10, 20, 40 μM)/miR-3657 inhibitor (0, 10, 20, 40 μM). (F) Schematic illustration of the sequences for STAT3 and the wild-type or mutated 3′-UTR of miR–3657, and dual Rluc/Fluc luciferase luminescence intensity of miR-3657 wild-type or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with STAT3 or pCDNA3.1. STAT3: signal transducer and activator of transcription 3. (G) qPCR to determine miR-3657 expression in hBMSCs treated with lenti-STAT3 and siR-STAT3 in a dose-dependent manner. n=3. Lenti-: lentivirus; siR-: siRNA; qPCR: quantitative polymerase chain reaction. (H) GLP-1R levels in hBMSCs after treatment with the STAT3 agonist and miR-3657 mimics (0, 10, 20, 40 μM) or treatment with the STAT3 inhibitor and miR-3657 inhibitor (0, 10, 20, 40 μM). (I) STAT3 levels in hBMSCs treated with metformin and the miR-140-3p inhibitor. (J) qPCR to determine miR-3657 expression in hBMSCs treated with metformin and miR-140–30 inhibitor/siR-STAT3. (K) Schematic illustration of the sequences for miR-140-3p and wild-type or mutated 3′-UTR of STAT3 mRNA, and dual Rluc/Fluc luciferase luminescence intensity of STAT3 wild-type or mutated 3′-UTR reporter plasmids in HEK293 cells co-transfected with miR-140-3p mimics or miR-NC mimics. (L, M) Protein expression of STAT3 in hBMSCs after treatment with miR-140-3p mimics (0, 10, 20, 40 μM) and with the miR-140-3p inhibitor (0, 10, 20, 40 μM).

Article Snippet: To figure out the effect of Sema4D on bone formation, mice were randomly divided into 5 groups (n = 6/group): control group, T2DM group, T2DM + Exendin-4 group (20 mg/kg, MCE,13241–33-3), T2DM + anti-Sema4D group (200 μg/kg, sc-390675), and T2DM + anti-Sema4D + Exendin-4 group.

Techniques: Expressing, Luciferase, Transfection, Real-time Polymerase Chain Reaction