Journal: EBioMedicine

Article Title: Reduced circulating BMP10 and BMP9 and elevated endoglin are associated with disease severity, decompensation and pulmonary vascular syndromes in patients with cirrhosis

doi: 10.1016/j.ebiom.2020.102794

Figure Lengend Snippet: BMP activity is low in liver disease but can be reconstituted with exogenous BMP9. (A,B) HAECs were serum-depleted overnight followed by addition of 1 ng/ml BMP9, BMP10, or 1% plasma from controls ( n = 6) or liver disease patients with undetectable (320; n = 5) levels of BMP9. After 1 h, cells were lysed, RNA extracted and cDNA analysed for the expression of (A) ID1 or (B) ID2 . (C and D) Plasmas (1% final concentration) from controls ( n = 4) or liver disease patients ( n = 4) were added alone, or after spiking with ProBMP9, to serum depleted HAECs for 1 h. After 1 h, cells were lysed, RNA extracted, and cDNA analysed for the expression of (C) ID1 or (D) ID2 . (E) The same spiked plasma samples ( n = 4 control and 4 liver disease) as in (C) and (D) were assayed using the BMP9 ELISA. * P< 0.05, ** P <0.01, *** P< 0.001 Kruskal Wallis test, # P< 0.05 Mann–Whitney test compared to control plasma.

Article Snippet: ELISA plates were coated with 0.5 μg/well of mouse monoclonal anti-human BMP10 antibody (MAB2926, R&D Systems) in PBS overnight at 4 °C.

Techniques: Activity Assay, Clinical Proteomics, Expressing, Concentration Assay, Control, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: EBioMedicine

Article Title: Reduced circulating BMP10 and BMP9 and elevated endoglin are associated with disease severity, decompensation and pulmonary vascular syndromes in patients with cirrhosis

doi: 10.1016/j.ebiom.2020.102794

Figure Lengend Snippet: Liver BMP9 and BMP10 are present in hepatocytes and their expression levels are reduced in cirrhosis, whereas endoglin expression is increased . (A) Low power image (10X) of control and cirrhotic liver sections stained with haematoxylin and eosin (White scale bar length = 30 µm). (B–D) Total RNA was extracted from control ( n = 8) and cirrhotic ( n = 9) liver samples and analysed by qPCR for (B) BMP9 , (C) BMP10 and (D) ENG expression. Mann Whitney Test * P< 0.05. (E,F) 8 healthy human liver samples and 9 samples from patients with cirrhosis were stained with Hoescht nuclear stain (blue) and a hepatocyte-specific antibody (red) in addition to antibodies (green) for (E) BMP9 or (F) BMP10. Images were captured using confocal microscopy and 100 µm scale bars are shown.(For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: ELISA plates were coated with 0.5 μg/well of mouse monoclonal anti-human BMP10 antibody (MAB2926, R&D Systems) in PBS overnight at 4 °C.

Techniques: Expressing, Control, Staining, MANN-WHITNEY, Confocal Microscopy

Journal: Clinical and Experimental Dental Research

Article Title: Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

doi: 10.1002/cre2.55

Figure Lengend Snippet: Growth factor adsorption of recombinant human bone morphogenetic protein 2 and recombinant human bone morphogenetic protein 9to absorbable collagen sponges at 0 min, 15 min, 60 min, 8 hrs, 24 hrs, 3 days, and 10 days as quantified by enzyme‐linked immunosorbent assay. absorbable collagen sponges were able to efficiently adsorb recombinant bone morphogenetic protein 2 and bone morphogenetic protein 9 and have comparable release kinetics over time

Article Snippet: Recombinant human BMP2 and rhBMP9 were purchased from R&D systems Inc (Minneapolis, MM, USA).

Techniques: Adsorption, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Experimental Dental Research

Article Title: Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

doi: 10.1002/cre2.55

Figure Lengend Snippet: (a) Attachment (8 hrs) and (b) proliferation (1, 3, and 5 days) assays of ST2 cells seeded on control tissue culture plastic, control absorbable collagen sponges (ACS), ACS loaded with bone morphogenetic protein 2 low (10 ng/ml), ACS loaded with bone morphogenetic protein 2 high (100 ng/ml), ACS loaded with bone morphogenetic protein 9 low (10 ng/ml), and ACS loaded with bone morphogenetic protein 9 high (100 ng/ml). No significant difference was observed between six groups at all time points

Article Snippet: Recombinant human BMP2 and rhBMP9 were purchased from R&D systems Inc (Minneapolis, MM, USA).

Techniques: Control

Journal: Clinical and Experimental Dental Research

Article Title: Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

doi: 10.1002/cre2.55

Figure Lengend Snippet: (a) Alkaline phosphatase staining of absorbable collagen sponges (ACS) at 7 days of ST2 cells seeded on control tissue culture plastic, control ACS, ACS loaded with BMP2 low (10 ng/ml), ACS loaded with BMP2 high (100 ng/ml), ACS loaded with BMP9 low (10 ng/ml), and ACS loaded with BMP9 high (100 ng/ml). (b) bone morphogenetic protein 9 low and high significantly increased alkaline phosphatase staining when compared to control and bone morphogenetic protein 2samples (* denotes significant difference, p < .05; ** denotes significantly higher than all other treatment modalities, p < .05)

Article Snippet: Recombinant human BMP2 and rhBMP9 were purchased from R&D systems Inc (Minneapolis, MM, USA).

Techniques: Staining, Control

Journal: Clinical and Experimental Dental Research

Article Title: Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

doi: 10.1002/cre2.55

Figure Lengend Snippet: Real‐time PCR of ST2 cells seeded on control tissue culture plastic, control absorbable collagen sponges (ACS), ACS loaded with BMP2 low (10 ng/ml), ACS loaded with BMP2 high (100 ng/ml), ACS loaded with BMP9 low (10 ng/ml), and ACS loaded with BMP9 high (100 ng/ml) for genes encoding (a) runt‐related transcription factor 2, (b) collagen 1 alpha 2 (COL1a2), (c) alkaline phosphatase (ALP), (d) bone sialoprotein (BSP), and (e) osteocalcin (OCN) at 3 and 14 days postseeding (* denotes significant difference, p < .05; ** denotes significantly higher than all other treatment modalities, p < .05)

Article Snippet: Recombinant human BMP2 and rhBMP9 were purchased from R&D systems Inc (Minneapolis, MM, USA).

Techniques: Real-time Polymerase Chain Reaction, Control

Journal: Clinical and Experimental Dental Research

Article Title: Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2

doi: 10.1002/cre2.55

Figure Lengend Snippet: (a) Visual representation of alizarin red‐stained of negative control absorbable collagen sponges (ACS) without cells, control tissue culture plastic, control ACS, ACS loaded with BMP2 low (10 ng/ml), ACS loaded with BMP2 high (100 ng/ml), ACS loaded with BMP9 low (10 ng/ml), and ACS loaded with BMP9 high (100 ng/ml) at 14 days postseeding. Note the intensity of red staining of ACS coated with BMP9 in comparison to control and BMP2 samples. (b) Quantified data of alizarin red staining from colour thresholding software (* denotes significant difference, p < .05; ** denotes significantly higher than all other treatment modalities, p < .05)

Article Snippet: Recombinant human BMP2 and rhBMP9 were purchased from R&D systems Inc (Minneapolis, MM, USA).

Techniques: Staining, Negative Control, Control, Comparison, Software

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Characterization of GDF2 Mutations and Levels of BMP9 and BMP10 in Pulmonary Arterial Hypertension

doi: 10.1164/rccm.201906-1141oc

Figure Lengend Snippet: Figure 1: Characterisation of Expressed BMP9 Mutant Proteins. (a) Schematic of

Article Snippet: After washing, 0.04 μg/well of anti-human BMP9 detection antibody (R&D Systems BAF3209) was added in Copyright © 2019 by the American Thoracic Society 6 PBS/1% BSA containing 0.2% GS.

Techniques: Mutagenesis

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Characterization of GDF2 Mutations and Levels of BMP9 and BMP10 in Pulmonary Arterial Hypertension

doi: 10.1164/rccm.201906-1141oc

Figure Lengend Snippet: Figure 2: Loss of Activity in Pro:BMP9 Mutants Predicted to be Pathogenic. (a,b)

Article Snippet: After washing, 0.04 μg/well of anti-human BMP9 detection antibody (R&D Systems BAF3209) was added in Copyright © 2019 by the American Thoracic Society 6 PBS/1% BSA containing 0.2% GS.

Techniques: Activity Assay

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Characterization of GDF2 Mutations and Levels of BMP9 and BMP10 in Pulmonary Arterial Hypertension

doi: 10.1164/rccm.201906-1141oc

Figure Lengend Snippet: Figure 3: Loss of active BMP9 in PAH patients carrying putatively pathogenic GDF2

Article Snippet: After washing, 0.04 μg/well of anti-human BMP9 detection antibody (R&D Systems BAF3209) was added in Copyright © 2019 by the American Thoracic Society 6 PBS/1% BSA containing 0.2% GS.

Techniques:

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Characterization of GDF2 Mutations and Levels of BMP9 and BMP10 in Pulmonary Arterial Hypertension

doi: 10.1164/rccm.201906-1141oc

Figure Lengend Snippet: Figure 4: Plasma BMP9 and pBMP10 levels are not reduced in PAH, but a subset of

Article Snippet: After washing, 0.04 μg/well of anti-human BMP9 detection antibody (R&D Systems BAF3209) was added in Copyright © 2019 by the American Thoracic Society 6 PBS/1% BSA containing 0.2% GS.

Techniques: Clinical Proteomics

Journal: Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein 1 Prodomain Specifically Binds and Regulates Signaling by Bone Morphogenetic Proteins 2 and 4

doi: 10.1074/jbc.m610929200

Figure Lengend Snippet: FIGURE 2. ProBMP1SSQQ binds BMP4 and BMP2 in a highly specific man- ner. A, an immunoblot is shown of samples containing () or lacking () 5 nM proBMP1SSQQ, BMP1, and/or BMP4; which were immunoprecipitated with anti-BMP1 antibodies (26). Blots were cut and the separate pieces were incu- bated with anti-BMP1 or anti-BMP4 antibodies. B, proBMP1SSQQ was incu- bated with a fixed amount of BMP4 (5 nM) alone, or in the presence of increas- ing amounts of BMP2 (molar ratios of BMP4:BMP2 of 1:1, 1:2, 1:5, and 1:10) followed by immunoprecipitation with anti-BMP1 antibody (26) and immu- noblotting with anti-BMP4 or anti-BMP2 monoclonal antibodies. As a control for pull-down efficiency, the anti-BMP4 blot was stripped and re-probed with anti-BMP1 antibodies to detect immunoprecipitated proBMP1SSQQ. C, puri- fied BMP1 prodomain, produced in a baculovirus system, was visualized by stainingwithCoomassieBrilliantBlueR-250(lane2).Numberstotheleftofgel correspond to the approximate sizes in kDa of molecular mass markers (lane 1). D, Western blot is used for characterization of anti-BMP prodomain anti- bodies, which at a 1:20,000 dilution detect 6 ng of BMP1-Fc fusion protein (lane 1), but do not detect 6 ng of Fc domain (lane 2).

Article Snippet: Recombinant BMP2, BMP4, BMP5, EGF, BMPR-1A/Fc, and TGF- 1 were all purchased from R&D systems.

Techniques: Western Blot, Immunoprecipitation, Bioprocessing, Control, Produced

Journal: Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein 1 Prodomain Specifically Binds and Regulates Signaling by Bone Morphogenetic Proteins 2 and 4

doi: 10.1074/jbc.m610929200

Figure Lengend Snippet: FIGURE 3. The BMP1 prodomain directly binds BMP2 and BMP4 with high specificity and affinity. A, 5 nM BMP4 and His-tagged BMP1 prodomain were incubated separately or together, followed by precipitation with nickel- chargedaffinityresin(leftpanel)orwithanti-BMP1prodomainantibody(right panel) and immunoblot analysis with anti-prodomain or anti-BMP4 antibod- ies.B,5nMBMP2andHis-taggedBMP1prodomainwereincubatedseparately or together, followed by precipitation with anti-BMP1 prodomain antibody and protein A-Sepharose, and immunoblot analysis with anti-BMP1 prodo- main or with anti-BMP2 antibodies. C, 5 nM BMP4 and His-tagged BMP1 prodomain were coincubated, as in A, but in the presence of 10-fold excesses ofBMP2,TGF-1,EGF,orBMP5;followedbyprecipitationwithnickel-charged affinity resin and immunoblot analyses. D, 5 nM BMP4 and soluble BMP recep- tor IA (BMPR-1A, ALK-3), fused to an Fc domain, were coincubated in the presence of 1:1, 2:1, or 5:1 molar ratios of either BMP1 prodomain or Chordin, and the samples then precipitated with protein A-Sepharose and analyzed by immunoblots employing anti-BMP4 or anti-BMPR-1A antibodies.

Article Snippet: Recombinant BMP2, BMP4, BMP5, EGF, BMPR-1A/Fc, and TGF- 1 were all purchased from R&D systems.

Techniques: Incubation, Western Blot

Journal: Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein 1 Prodomain Specifically Binds and Regulates Signaling by Bone Morphogenetic Proteins 2 and 4

doi: 10.1074/jbc.m610929200

Figure Lengend Snippet: FIGURE 4. The BMP1 prodomain binds BMP2 with a KD of 10.9 nM. Coomassie Brilliant Blue R-250-stained SDS-PAGE gels are shown for molecular mass standards (lane 1) and purified BMP1 prodomain-Fc fusion protein (lane 2) (A) or molecular mass standards (lane 1) and purified BMP1 prodomain minus the Fc domain (lane 2) (B). Numbers to the left of gels correspond to the approximate sizes in kDa of molecular mass markers. BMP1 prodomain (C) and murine Chordin (D) bind to BMP2 with KDs of 10.9 4.7 nM and 6.7 1.0 nM, respectively.

Article Snippet: Recombinant BMP2, BMP4, BMP5, EGF, BMPR-1A/Fc, and TGF- 1 were all purchased from R&D systems.

Techniques: Staining, SDS Page, Purification

Journal: Immunity

Article Title: Stellate Cells, Hepatocytes, and Endothelial Cells Imprint the Kupffer Cell Identity on Monocytes Colonizing the Liver Macrophage Niche

doi: 10.1016/j.immuni.2019.08.017

Figure Lengend Snippet:

Article Snippet: Recombinant Human/Mouse/Rat BMP-9 Protein , RnD Systems , Cat#5566-BP-010.

Techniques: Control, Recombinant, Blocking Assay, Irradiation, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Microarray, Software, Microscopy