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The osteogenic signal transduction of OP5. (A) The binding affinity of the peptide to BMP receptor types IA and II <t>(BMPR-IA</t> and II). BMPR-IA or BMPR-II in lysate of human mesenchymal stromal cells (hMSCs) was immobilized on a microplate using a specific BMP receptor antibody in an ELISA. Data represents mean ± S.D. Binding activity of BMP-2 (2 μg/ml) and OP5 (20 μg/ml) are detected using anti-BMP-2 monoclonal antibody <t>and</t> <t>1:2000</t> horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG as a secondary antibody binding to primary antibody. Each bar shows the mean ± S.D. ∗ P < 0.05, ∗∗ P < 0.01. (B) Western blot analysis of β-catenin, phosphorylated cAMP response element-binding protein (P-CREB), and phosphorylated SMAD (P-SMAD) expression in hMSCs treated with BMP-2 and OP5 for 3 and 6 h. Cell lysates were prepared and analyzed by western blot using antibodies to β-catenin, P-CREB, and P-SMAD (all of 1:1000). The expression level of each group was normalized with the density % of β-actin in the same group. O.D., optical density; None, without peptide or protein group; BMP, bone morphogenetic protein; OP, osteogenic peptide; DWIVA, amino acid sequence from osteogenic peptide; 10F, 10F medium [Low-glucose Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum (FBS) and 1 % antibiotic/antimycotic] as the control group.
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The osteogenic signal transduction of OP5. (A) The binding affinity of the peptide to BMP receptor types IA and II (BMPR-IA and II). BMPR-IA or BMPR-II in lysate of human mesenchymal stromal cells (hMSCs) was immobilized on a microplate using a specific BMP receptor antibody in an ELISA. Data represents mean ± S.D. Binding activity of BMP-2 (2 μg/ml) and OP5 (20 μg/ml) are detected using anti-BMP-2 monoclonal antibody and 1:2000 horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG as a secondary antibody binding to primary antibody. Each bar shows the mean ± S.D. ∗ P < 0.05, ∗∗ P < 0.01. (B) Western blot analysis of β-catenin, phosphorylated cAMP response element-binding protein (P-CREB), and phosphorylated SMAD (P-SMAD) expression in hMSCs treated with BMP-2 and OP5 for 3 and 6 h. Cell lysates were prepared and analyzed by western blot using antibodies to β-catenin, P-CREB, and P-SMAD (all of 1:1000). The expression level of each group was normalized with the density % of β-actin in the same group. O.D., optical density; None, without peptide or protein group; BMP, bone morphogenetic protein; OP, osteogenic peptide; DWIVA, amino acid sequence from osteogenic peptide; 10F, 10F medium [Low-glucose Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum (FBS) and 1 % antibiotic/antimycotic] as the control group.

Journal: Regenerative Therapy

Article Title: Bone morphogenetic protein-2-derived osteogenic peptide promotes bone regeneration via osteoblastogenesis

doi: 10.1016/j.reth.2025.09.006

Figure Lengend Snippet: The osteogenic signal transduction of OP5. (A) The binding affinity of the peptide to BMP receptor types IA and II (BMPR-IA and II). BMPR-IA or BMPR-II in lysate of human mesenchymal stromal cells (hMSCs) was immobilized on a microplate using a specific BMP receptor antibody in an ELISA. Data represents mean ± S.D. Binding activity of BMP-2 (2 μg/ml) and OP5 (20 μg/ml) are detected using anti-BMP-2 monoclonal antibody and 1:2000 horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgG as a secondary antibody binding to primary antibody. Each bar shows the mean ± S.D. ∗ P < 0.05, ∗∗ P < 0.01. (B) Western blot analysis of β-catenin, phosphorylated cAMP response element-binding protein (P-CREB), and phosphorylated SMAD (P-SMAD) expression in hMSCs treated with BMP-2 and OP5 for 3 and 6 h. Cell lysates were prepared and analyzed by western blot using antibodies to β-catenin, P-CREB, and P-SMAD (all of 1:1000). The expression level of each group was normalized with the density % of β-actin in the same group. O.D., optical density; None, without peptide or protein group; BMP, bone morphogenetic protein; OP, osteogenic peptide; DWIVA, amino acid sequence from osteogenic peptide; 10F, 10F medium [Low-glucose Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum (FBS) and 1 % antibiotic/antimycotic] as the control group.

Article Snippet: The plates were then incubated overnight at 4 °C with 1:1,000 mouse anti-BMPR-IA and anti-BMPR-II antibody (R&D systems), followed by reaction with 1:2000 horseradish peroxidase (HRP)-conjugate rabbit anti-mouse IgG (catalog no. 7076; Cell Signaling Technology Inc., Danvers, MA, USA) for 30 min at room temperature.

Techniques: Transduction, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Western Blot, Expressing, Sequencing, Modification, Control