recombinant human gdf15 protein  (R&D Systems)

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: Dysregulation of BMP9/ALK1 signaling and inflammation in refractory ulcerative colitis (rUC). ( A ) Heatmap depicting serum expression levels of BMP family members in healthy controls, non-rUC, and rUC patients ( n = 3 per group). Data were Z-score normalized and hierarchically clustered (red, high expression; blue, low expression). ( B ) Baseline serum levels of BMP9 and BMP10 in healthy controls ( n = 50), non-rUC patients ( n = 47), and rUC patients ( n = 48). ( C ) Spearman correlation analyses between baseline serum BMP9 levels and clinical disease activity indices, including baseline Modified Mayo Score, baseline UCEIS, post-treatment Modified Mayo Score, and post-treatment UCEIS, in patients with UC ( n = 95). ( D ) Colonic mucosal mRNA expression levels of ALK1, IL-6, TNF-α, and CCL2 in healthy controls, non-rUC, and rUC patients ( n = 4 per group). Statistical annotations for ( B , D ): (Normalized to GAPDH; Mean ± SD; Statistical significance determined by one-way ANOVA with Tukey’s post hoc test: ** p < 0.01, *** p < 0.001, ns: not significant).

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Expressing, Activity Assay, Modification

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 attenuates DSS-induced colitis in mice. ( A ) Schematic of experimental design: Acute colitis was induced in C57BL/6 mice by 3% DSS in drinking water for 7 days. The BMP9 treatment group received intraperitoneal injections of recombinant murine BMP9 (200 ng/day), while the DSS group and the control group received PBS ( n = 14 per group). ( B ) Serum BMP9(ng/mL) concentrations measured by ELISA. ( C ) Representative images of colons and quantitative analysis of colon length (cm). ( D ) Colonoscopy images (upper), H&E-stained colon sections (lower; scale bars = 100 μm), and histopathological scores. ( E ) (Upper) Dynamic body weight changes and (Lower) Disease Activity Index (DAI) scores. Data expressed as mean ± SD; * p < 0.05, two-way repeated measures ANOVA. ( F ) (Left) Relative Alk1 mRNA expression in colon tissues (RT-qPCR normalized to Gapdh). (Right) ALK1 protein concentrations (quantified by ELISA). Data presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 ns: not significant). ( G ) Relative mRNA expression levels of IL-1β, Ccl2, Col1a1, and Col3a1 (RT-qPCR; mean ± SD). ( H ) Western blot analysis of CCL2, TGF-β, and α-SMA protein expression in colon tissues and quantitative analysis of band intensities. ( I ) Western blot detection of p-Smad1, total Smad1, and VE-cadherin proteins and quantitative analysis of band intensities.

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Recombinant, Control, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 restores intestinal vascular barrier integrity in DSS-induced colitis. ( A ) Representative immunofluorescence images of VE-cadherin (red) and CD31 (green) co-localization in colon tissues (nuclei counterstained with DAPI). ( B ) Left: Schematic of FITC-dextran (4 kDa) permeability assay. Right: Quantified serum FITC fluorescence intensity 60 min post-gavage ( n = 5 per group). ( C ) Left: Schematic of Evans Blue vascular leakage assay. Right: Colonic Evans Blue extravasation quantified by absorbance at 620 nm (mean ± SD; n = 5 per group; ** p < 0.01, *** p < 0.001, ns: not significant; one-way ANOVA with Tukey’s test). ( D ) KEGG pathway analysis of RNA-seq data from colon tissues (DSS + BMP9 groups vs. DSS). ( E ) Volcano plot of differentially expressed genes. Genes highlighted in bold are key IBD-associated downregulated factors.

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Immunofluorescence, Permeability, Fluorescence, RNA Sequencing

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9/ALK1 signaling modulates neutrophil migration and endothelial tube formation. ( A ) Schematic representation of the neutrophil migration assay. ( B ) (Left) Representative fluorescence micrographs demonstrating Calcein-AM-labeled neutrophil migration through 3 μm pore Transwell inserts toward HIMECs pretreated for 2 h with: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL as positive control), or ALK1 inhibitor ML347 (150 nM). (Right) Quantitative analysis of neutrophil migration rates (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant by two-way ANOVA with post hoc testing). ( C ) Schematic illustration of the endothelial tube formation assay protocol. ( D ) Representative phase-contrast images of tubular network formation by HIMECs cultured on growth factor-reduced Matrigel under various treatment conditions: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL), or ML347 (150 nM). ( E ) Quantitative assessment of angiogenic parameters including branch points and nodal junctions (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA with appropriate post hoc comparisons).

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Migration, Fluorescence, Labeling, Control, Positive Control, Endothelial Tube Formation Assay, Cell Culture

Journal: Journal of Orthopaedic Translation

Article Title: Asymmetrical paravertebral muscles fibrosis causes progression of adolescent idiopathic scoliosis via myostatin signalling in fibro-adipogenic progenitors

doi: 10.1016/j.jot.2025.11.003

Figure Lengend Snippet: Asymmetrical myostatin activation of fibro-adipogenic progenitors (FAPs) in PVMs (A) The UMAP plots of main cell types for the PVMs. FAPs = fibro-adipogenic progenitors; SMCs = smooth muscle cells; MuSCs = muscle stem cells. (B) The UMAP plots of main cell types for the PVMs from the concave and convex side. (C) Dot plot of highly expressed genes in each cell type. (D) Percentage of each cell type. (E) UMAP plots of genes marked FAPs. (F) Pathway enrichment analysis of differentially expressed genes in FAPs. (G) Hierarchical clustering heatmap illustrating differentially expressed genes (DEGs) between the concave and convex side of PVMs, with MSTN (myostatin) marked (red arrow). (H)Volcano plot illustrating DEGs between the concave and convex side of the PVMs. (I) Co-localization of Laminin (red), PDGFRα (green), and Myostatin (yellow) on the concave and convex side of the PVMs in patients with AIS. Scale bar: 50 μm. (J) Quantification of data I, n = 5. Paired t-tests, ∗∗p < 0.01.

Article Snippet: Mice were randomly categorized into two groups (n = 7 per group) after surgery: (1) the Unilateral Myostatin group in which 100 nM recombinant myostatin (80 μL, HY- P72632 , MCE) was administered by injection into the left PVMs twice weekly for 4 weeks, with 80 μL 0.1 % dimethyl sulfoxide (DMSO, ST038, Beyotime) was injected contralaterally. (2) the Bilateral DMSO group in which 80 μL 0.1 % DMSO was bilaterally injected into the PVMs.

Techniques: Activation Assay

Journal: Journal of Orthopaedic Translation

Article Title: Asymmetrical paravertebral muscles fibrosis causes progression of adolescent idiopathic scoliosis via myostatin signalling in fibro-adipogenic progenitors

doi: 10.1016/j.jot.2025.11.003

Figure Lengend Snippet: Myostatin promotes fibro-differentiation of FAPs in vitro . (A) Flow cytometry analysis for isolating FAPs from mouse skeletal muscle using fluorescence-activated cell sorting based on PDGFRα, CD31, CD45, and integrin-α7 expression. The P4 gates correspond to FAPs populations. (B) Immunofluorescence staining for ki-67 (red) in FAPs with or without 100 nM myostatin treatment for 2 days. Scale bar: 50 μm. (C) Quantification of data B. n = 3. Unpaired t-tests, ∗∗p < 0.01. (D) Oil Red O staining (red) and immunofluorescence staining for Perilipin (green) in FAPs cultured in differentiation medium with or without dexamethasone (Dex) or myostatin for 1 week. Scale bar: 50 μm. (E) Quantification of data D. n = 3. One-way ANOVA with Tukey's post-hoc test, ns = no significant; ∗∗∗p < 0.001. (F) Immunofluorescence staining for α-SMA (red) in FAPs cultured in differentiation medium with or without Dex or myostatin for 1 week. Scale bar: 50 μm. (G) Quantification of data F, n = 3. One-way ANOVA with Tukey's post-hoc test, ns = no significant; ∗∗p < 0.01. (H) Western blot analysis of α-SMA and COL1 in FAPs treated with Dex or myostatin for 1 week. (I) Quantification of data H, n = 3. One-way ANOVA with Tukey's post-hoc test, ns = no significant; ∗∗∗p < 0.001. (J) qPCR analysis of fibrotic gene expression ( Col1a1, Col6a1, Acta2 ) in FAPs after 2 days of Dex or myostatin treatment. n = 3. One-way ANOVA with Tukey's post-hoc test, ns = not significant; ∗∗∗p < 0.001.

Article Snippet: Mice were randomly categorized into two groups (n = 7 per group) after surgery: (1) the Unilateral Myostatin group in which 100 nM recombinant myostatin (80 μL, HY- P72632 , MCE) was administered by injection into the left PVMs twice weekly for 4 weeks, with 80 μL 0.1 % dimethyl sulfoxide (DMSO, ST038, Beyotime) was injected contralaterally. (2) the Bilateral DMSO group in which 80 μL 0.1 % DMSO was bilaterally injected into the PVMs.

Techniques: In Vitro, Flow Cytometry, Fluorescence, FACS, Expressing, Immunofluorescence, Staining, Cell Culture, Western Blot, Gene Expression

Journal: Journal of Orthopaedic Translation

Article Title: Asymmetrical paravertebral muscles fibrosis causes progression of adolescent idiopathic scoliosis via myostatin signalling in fibro-adipogenic progenitors

doi: 10.1016/j.jot.2025.11.003

Figure Lengend Snippet: Asymmetric activation of myostatin in the PVMs leads to scoliosis in a bipedal mouse model. (A) Schematic of the experimental design. Three-week-old bipedal female mice received either bilateral injections of dimethyl sulfoxide (DMSO) (Bilateral DMSO group) or injections of recombinant myostatin (left side) and DMSO (right side) (Unilateral Myostatin group) into the PVMs, administered twice weekly for 4 weeks. Spine evaluation and tissue harvested were performed 2 weeks after the final injection. (B) Representative X-ray and micro-CT images in the bilateral DMSO group and the unilateral myostatin group. (C) Representative H&E and Sirius Red staining of the PVMs in the bilateral DMSO group and the unilateral myostatin group. Scale bar: 200 μm. (D) Quantification of coronal spinal curvature in the two groups. n = 7. Unpaired t-tests, ∗∗∗p < 0.001. (E) Quantification of sagittal spinal curvature in the two groups. n = 7. Unpaired t-tests, ∗∗P < 0.01. (F) Quantification of fibrotic area based on Sirius Red staining, shown in data (C) above. n = 7. One-way ANOVA with Tukey's post-hoc test, ∗∗P < 0.01. (G) Western blot analysis of COL1 and α-SMA in the left and right side of the PVMs of the Unilateral Myostatin group. (H) Laminin (red) and COL1 (green) co-immunofluorescence in the left and right side of PVMs in the Unilateral Myostatin group. Scale bar: 100 μm. (I) Quantification of data G, n = 7. Paired t-tests, ∗∗p < 0.01. (J) qPCR analysis of Col1a1, Col6a1 and Acta2 in the left and right side of the PVMs of the two groups. n = 7. One-way ANOVA with Tukey's post-hoc test, ns = no significant; ∗P < 0.05; ∗∗∗p < 0.001.

Article Snippet: Mice were randomly categorized into two groups (n = 7 per group) after surgery: (1) the Unilateral Myostatin group in which 100 nM recombinant myostatin (80 μL, HY- P72632 , MCE) was administered by injection into the left PVMs twice weekly for 4 weeks, with 80 μL 0.1 % dimethyl sulfoxide (DMSO, ST038, Beyotime) was injected contralaterally. (2) the Bilateral DMSO group in which 80 μL 0.1 % DMSO was bilaterally injected into the PVMs.

Techniques: Activation Assay, Recombinant, Injection, Micro-CT, Staining, Western Blot, Immunofluorescence

Journal: Journal of Orthopaedic Translation

Article Title: Asymmetrical paravertebral muscles fibrosis causes progression of adolescent idiopathic scoliosis via myostatin signalling in fibro-adipogenic progenitors

doi: 10.1016/j.jot.2025.11.003

Figure Lengend Snippet: Myostatin promotes FAPs differentiation into fibrocytes via SMAD3 activation. (A) Myostatin (yellow) and p-SMAD3 (cyan) co-immunofluorescence in the concave and convex sides of PVMs from AIS Patients. Scale bars: 200 μm. Right: Quantification of Myostatin and p-SMAD3 positive cells. n = 10. Paired t-tests, ∗∗∗p < 0.001. (B) Western blot analysis of SMAD3 and p-SMAD3 in FAPs treated without or with 100 nM recombinant myostatin for 48 h. (C) Quantitation of data B, n = 3. Unpaired t-tests, ns = no significant; ∗∗∗p < 0.001. (D) Western blot analysis of SMAD3, p-SMAD3, COL1, aSMA in FAPs transfected with control siRNA or siRNA-Smad3, with or without myostatin treatment for 1 week. (E) Immunofluorescence staining for aSMA in FAPs transfected with control siRNA or siRNA-Smad3, with or without myostatin treatment for 1 week. (F) Quantification of data D, n = 3. One-way ANOVA with Tukey's post-hoc test, ns = no significant; ∗∗P < 0.01; ∗∗∗p < 0.001. (G) Western blot analysis of SMAD3, p-SMAD3, COL1 and aSMA in FAPs transfected with overexpress plasmid-Smad3 with or not with myostatin for 1 week. (H) Quantification of data G, n = 3. One-way ANOVA with Tukey's post-hoc test, ∗p < 0.05; ∗∗p < 0.01;∗∗p < 0.001.

Article Snippet: Mice were randomly categorized into two groups (n = 7 per group) after surgery: (1) the Unilateral Myostatin group in which 100 nM recombinant myostatin (80 μL, HY- P72632 , MCE) was administered by injection into the left PVMs twice weekly for 4 weeks, with 80 μL 0.1 % dimethyl sulfoxide (DMSO, ST038, Beyotime) was injected contralaterally. (2) the Bilateral DMSO group in which 80 μL 0.1 % DMSO was bilaterally injected into the PVMs.

Techniques: Activation Assay, Immunofluorescence, Western Blot, Recombinant, Quantitation Assay, Transfection, Control, Staining, Plasmid Preparation

Journal: Journal of Orthopaedic Translation

Article Title: Asymmetrical paravertebral muscles fibrosis causes progression of adolescent idiopathic scoliosis via myostatin signalling in fibro-adipogenic progenitors

doi: 10.1016/j.jot.2025.11.003

Figure Lengend Snippet: Inhibition of myostatin–SMAD3 signalling attenuates paravertebral muscles (PVMs) fibrosis and scoliosis progression in a mouse model (A): Schematic of the experimental design. Three-week-old bipedal female mice received left-side injections of recombinant myostatin and right-side injections of DMSO into the PVMs twice weekly for 4 weeks. Mice were randomly assigned to three groups (n = 6 per group) after 2 weeks: the myostatin inhibitory protein treatment group (MIP group), the SMAD3 inhibitor small molecule 3 treatment group (SIS3 group), and the control group. The MIP/SIS3 group received injections of MIP or SIS3 into the left side of the PVMs every 3.5 days for 2 weeks. The control group received bilateral injections of DMSO into the PVMs. Spine evaluation and tissue harvesting were conducted 2 weeks after the final injection. (B) Representative HE and Sirius Red staining of the left side PVMs in the control group, MIP group, and SIS3 group. Scale bar: 200 μm. (C) Quantification of fibrotic area based on Sirius Red staining in data (B) above. n = 6. One-way ANOVA with Tukey's post-hoc test, ns = no significant; ∗∗∗P < 0.001. (D) Laminin (red) and COL1 (green) co-immunofluorescence in the left side PVMs in the control group, MIP group and SIS3 group. Scale bar: 100 μm. (E) Representative X-ray images in the control group, MIP group and SIS3 group at 6 weeks and 10 weeks. (F) Quantification of the coronal and sagittal spinal curvature in the three groups at 6 weeks and 10 weeks. n = 6. One-way ANOVA with Tukey's post-hoc test, ns = not significant; ∗P < 0.05; ∗∗P < 0.01.

Article Snippet: Mice were randomly categorized into two groups (n = 7 per group) after surgery: (1) the Unilateral Myostatin group in which 100 nM recombinant myostatin (80 μL, HY- P72632 , MCE) was administered by injection into the left PVMs twice weekly for 4 weeks, with 80 μL 0.1 % dimethyl sulfoxide (DMSO, ST038, Beyotime) was injected contralaterally. (2) the Bilateral DMSO group in which 80 μL 0.1 % DMSO was bilaterally injected into the PVMs.

Techniques: Inhibition, Muscles, Recombinant, Control, Injection, Staining, Immunofluorescence

Journal: Journal of Orthopaedic Translation

Article Title: Asymmetrical paravertebral muscles fibrosis causes progression of adolescent idiopathic scoliosis via myostatin signalling in fibro-adipogenic progenitors

doi: 10.1016/j.jot.2025.11.003

Figure Lengend Snippet: Schematic diagram of asymmetrical myostatin-SMAD3 activation in FAPs results in paravertebral muscle fibrosis and progression of AIS.

Article Snippet: Mice were randomly categorized into two groups (n = 7 per group) after surgery: (1) the Unilateral Myostatin group in which 100 nM recombinant myostatin (80 μL, HY- P72632 , MCE) was administered by injection into the left PVMs twice weekly for 4 weeks, with 80 μL 0.1 % dimethyl sulfoxide (DMSO, ST038, Beyotime) was injected contralaterally. (2) the Bilateral DMSO group in which 80 μL 0.1 % DMSO was bilaterally injected into the PVMs.

Techniques: Activation Assay