bmpr ii  (R&D Systems)

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

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis.

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Figure 5. Characterization of the interaction between GDF10 and BMPR2/ALK3. A) Percentage of expression+ binding+ transfectants of HEK293 cells expressing a single TGF-𝛽superfamily receptor incubated with GDF10-Fc. B) Percentage of expression+ binding+ transfectants of HEK293 cells express- ing BMPR2 and type I TGF-𝛽receptor incubated with GDF10-Fc. C) HEK293 cells were transfected to express BMPR2 and ALK3 and then incubated with GDF10-Fc, scale bars, 50 μm. D) Association of BMPR2 with ALK3. HEK293 cells were transfected to express BMPR2-FLAG and ALK3-HIS. E) The overall structure of mature GDF10 with the extracellular domains (ECD) of mouse BMPR2 and ALK3 (GDF10: blue-purple, BMPR2: pink-green, ALK3: pink). F) Western blot analysis of cell lysates from HEK293 cells transfected to express C-terminal FLAG-tagged BMPR2 and FLAG-tagged ALK3 incubated with GDF10-Fc and then immunoprecipitated with anti-FLAG antibody. G) Biacore sensorgrams and binding kinetics were determined by SPR spectroscopy for GDF10 with the ECD of mouse BMPR2 and ALK3. H) qPCR analysis of the expression Bmpr2 and Alk3 in mouse LSECs, MACs, HSCs, and HCs (n = 6). I) Cell type enrichment of BMPR2 and ALK3 in human liver (Human Protein Atlas database).

Article Snippet: For Western blot analysis, the following primary antibodies were employed: β-Actin (AC026; ABclonal), GDF10 (ab235005; Abcam), ACTA2 (A17910; ABclonal), COL1A1 (A22089; ABclonal), phospho-SMAD2-S467 (AP0269; ABclonal), SMAD2 (A19114; ABclonal), phospho-SMAD3-S423/S425 (AP1263; ABclonal), SMAD3 (A19115; ABclonal), phospho-SMAD1/5/8 (#13 820; Cell Signaling Technology), SMAD15/8 (A17439; ABclonal), BMPR2 (sc-393304; Santa Cruz Biotechnology), ALK3 (sc-134285; Santa Cruz Biotechnology), SMAD7 (sc-365846; Santa Cruz Biotechnology).

Techniques: Expressing, Binding Assay, Incubation, Transfection, Western Blot, Immunoprecipitation, Spectroscopy

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis.

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Figure 6. GDF10 inhibits HSC activation via the BMPR2/ALK3-SMAD1/5/8-SMAD7 signaling pathway. A,B) qPCR (A) (n = 6) and Western blot (B) analysis of indicated genes in the primary mouse HSCs infected with LV-sh-Luc or LV-sh-Bmpr2 and LV-sh-Alk3 for 24 h and then treated with TGF-𝛽1 plus Fc or GDF10-Fc for another 24 h. C) IF staining analysis of ACTA2 expression in HSCs treated as in (A), scale bars, 20 μm (n = 3). D, E) qPCR (n = 6) (D) and Western blot (E) analysis of indicated genes in the HSCs treated with TGF-𝛽1 plus GDF10-Fc and/or LDN for 24 h. F) IF analysis of ACTA2 stating in HSCs treated as in (D). G) Heat map shows the indicated genes in JS1 cells treated with TGF-𝛽1 plus LDN and/or GDF10-Fc for 24 h.

Article Snippet: For Western blot analysis, the following primary antibodies were employed: β-Actin (AC026; ABclonal), GDF10 (ab235005; Abcam), ACTA2 (A17910; ABclonal), COL1A1 (A22089; ABclonal), phospho-SMAD2-S467 (AP0269; ABclonal), SMAD2 (A19114; ABclonal), phospho-SMAD3-S423/S425 (AP1263; ABclonal), SMAD3 (A19115; ABclonal), phospho-SMAD1/5/8 (#13 820; Cell Signaling Technology), SMAD15/8 (A17439; ABclonal), BMPR2 (sc-393304; Santa Cruz Biotechnology), ALK3 (sc-134285; Santa Cruz Biotechnology), SMAD7 (sc-365846; Santa Cruz Biotechnology).

Techniques: Activation Assay, Western Blot, Infection, Staining, Expressing

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Autocrine GDF10 Inhibits Hepatic Stellate Cell Activation via BMPR2/ALK3 Receptor to Prevent Liver Fibrosis.

doi: 10.1002/advs.202500616

Figure Lengend Snippet: Figure 7. BMPR2:ALK3-Fc prevents the antifibrotic effect of GDF10 in the liver. A) Schematic drawing of the experimental procedure in 3-month-old male LoxP and Gdf10TG mice. B) Representative images of H&E, Sirius Red, and ACTA2 IHC staining, scale bars, 50 μm (n = 3). C) Hydroxyproline assay analysis of the total liver collagen of mice treated as in (A) (n = 6). D,E) qPCR (D) (n = 6) and Western blot (E) analysis of indicated genes in the liver of mice treated as in (A). F,G) qPCR (F) (n = 6) and Western blot (G) analysis of indicated genes in the HSCs from mice treated as in (A) (n = 3). H,I) IF staining (H) and Western blot (I) analysis of indicated protein in the HSCs from mice treated as in (A), scale bars, 5 μm. Data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by the one-way ANOVA (C,E,G,I) or two-way ANOVA (D,F).

Article Snippet: For Western blot analysis, the following primary antibodies were employed: β-Actin (AC026; ABclonal), GDF10 (ab235005; Abcam), ACTA2 (A17910; ABclonal), COL1A1 (A22089; ABclonal), phospho-SMAD2-S467 (AP0269; ABclonal), SMAD2 (A19114; ABclonal), phospho-SMAD3-S423/S425 (AP1263; ABclonal), SMAD3 (A19115; ABclonal), phospho-SMAD1/5/8 (#13 820; Cell Signaling Technology), SMAD15/8 (A17439; ABclonal), BMPR2 (sc-393304; Santa Cruz Biotechnology), ALK3 (sc-134285; Santa Cruz Biotechnology), SMAD7 (sc-365846; Santa Cruz Biotechnology).

Techniques: Immunohistochemistry, Hydroxyproline Assay, Western Blot, Staining