bmp10  (R&D Systems)

Journal: Angiogenesis

Article Title: Genetic and pharmacological targeting of mTORC1 in mouse models of arteriovenous malformation expose non-cell autonomous signalling in HHT

doi: 10.1007/s10456-024-09961-5

Figure Lengend Snippet: EC-specific overactivation of mTORC1 reduces incidence and thickness of AVMs induced by blocking antibodies. A . Time scheme for administration of tamoxifen, BMP9/10 blocking antibodies (bAbs)/Isotype IgG, and sample collection. B . Representative pictures showing retinal vasculatures of control (Cre-) and Tsc1 iECKO mice treated with IgGs or anti-BMP9/10 antibodies (Abs). Arrows indicate AVMs. C . Quantification of number of AVMs per retina of Control ( n = 5 mice) and Tsc1 iECKO ( n = 5 mice) mice treated with anti-BMP9/10 Abs. D . Quantification of AVM thickness in Control ( n = 5 mice) and Tsc1 iECKO mice ( n = 3) treated with IgGs or anti-BMP9/10 Abs. E . Central retinal vasculatures of control (Cre-) and Tsc1 iECKO mice, treated with anti-BMP9/10 Abs, immunolabelled for CD31, p-RPS6, and YFP (indicative of recombination). F . Quantification of the ratio of the p-RPS6 + area within the CD31 + area to the complete CD31 + area ( F ), as well as the p-RPS6 + area outside the CD31 + area to the complete non-vascular area ( G ) of central retinas of Control ( n = 3 mice) and Tsc1 iECKO ( n = 3 mice) mice treated with anti-BMP9/10 Abs. Note the vascular specific increase of p-RPS6, indicative of EC-specific mTORC1 overactivation. All data was analysed by two-tailed unpaired t-test with Welch’s correction. Bars indicate mean ± s.d. * p < 0.05

Article Snippet: Pups were treated with mouse monoclonal antibodies against BMP9 and BMP10 (15 mg/kg, IgG2b, MAB3209; 15 mg/kg, IgG2a, MAB2926; R&D Systems, respectively) or isotype control antibodies (15 mg/kg, IgG2b, MAB004; 15 mg/kg, IgG2a, MAB003; R&D Systems, respectively) at P3, P4 and P5 and tissues were harvested at P6.

Techniques: Blocking Assay, Control, Two Tailed Test

Journal: EMBO Molecular Medicine

Article Title: BMP-9 mediates fibroproliferation in fibrodysplasia ossificans progressiva through TGF-β signaling

doi: 10.1038/s44321-024-00174-3

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Human/Mouse/Primate BMP-9 monoclonal antibody , R&D Systems , MAB3209.

Techniques: Transgenic Assay, Recombinant, Cell Counting, Cell Cycle Assay, Binding Assay, Blocking Assay, Enzyme-linked Immunosorbent Assay, Luciferase, Reporter Assay, Western Blot, Reverse Transcription, Control, Software, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: CDK6-mediated endothelial cell cycle acceleration drives arteriovenous malformations in hereditary hemorrhagic telangiectasia

doi: 10.1101/2023.09.15.554413

Figure Lengend Snippet: ( A ) Schematic representation of the i.p. injection protocol. Arrowheads indicate the postnatal days (P) of injection (AM or PM). Pups were euthanized at P6 for analysis. ( B ) Representative staining using isolectin B4 (IB4, green), and of ERG (blue) and EdU (red) in the peri-optic nerve and mid-plexus regions of retinas from PBS controls, vehicle-treated BMP9/10ib mice, and palbociclib-treated BMP9/10ib mice. Arrows denote AVMs; a, artery; v, vein. Scale bars, 50 μm. ( C and D ) Scatter plots showing the total number of ERG + cells per 200 μm in arteries ( C ) and veins ( D ) in the peri-optic nerve region across three groups: PBS (n=11), vehicle-treated BMP9/10ib (n=9), and palbociclib-treated BMP9/10ib (n=12) mice. ( E and F ) Scatter plots showing ERG + EdU + cells in the peri-optic nerve ( E ) and mid-plexus ( F ) regions across three groups: PBS (n=4-6), vehicle-treated BMP9/10ib (n=4-6), and palbociclib-treated BMP9/10ib (n=6) mice. Data represent individual retinas and mean ± SEM, one-way ANOVA with Tukey’s multiple comparisons test. ns, not significant; *P < 0.05, ***P < 0.001, ****P < 0.0001.

Article Snippet: BMP9 (25 mg/kg; MAB3209, R&D System) and BMP10 (50 mg/kg; MAB2926, R&D System) were injected intraperitoneally (i.p.) at P3 and P4.

Techniques: Injection, Staining

Journal: bioRxiv

Article Title: CDK6-mediated endothelial cell cycle acceleration drives arteriovenous malformations in hereditary hemorrhagic telangiectasia

doi: 10.1101/2023.09.15.554413

Figure Lengend Snippet: ( A and B ) Representative flow cytometry plots ( A ) and quantification ( B ) of cell cycle phase distribution of ECs isolated from livers of P6 and P8 PBS (n=14) and BMP9/10ib (n=11) mice. ( C and D ) Representative flow cytometry ( C ) and quantification ( D ) of cells analyzed as in ( A ) showing S phase duration measured as intra-S phase of EdU incorporation [intra-S EdU mean fluorescence intensity (MFI)]. Data in ( B ) and ( D ) represent mean ± SEM, unpaired t-test. ( E ) Schematic representation of the i.p. injection protocol. Arrowheads indicate the postnatal days (P) of injection (AM or PM). Pups were euthanized at P8 for analysis. ( F and G ) Representative flow cytometry plots of cell cycle speed analysis in iH2B-FT ECs ( F ) and quantification of fast-cycling iH2B-FT ECs ( G ) isolated from P8 livers of PBS (n=7), vehicle-treated BMP9/10ib (n=8), and palbociclib-treated BMP9/10ib (n=7) mice. Data in ( G ) represent mean ± SEM, one-way ANOVA with Tukey’s multiple comparisons test. ns, not significant; **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: BMP9 (25 mg/kg; MAB3209, R&D System) and BMP10 (50 mg/kg; MAB2926, R&D System) were injected intraperitoneally (i.p.) at P3 and P4.

Techniques: Flow Cytometry, Isolation, Fluorescence, Injection

Journal: bioRxiv

Article Title: CDK6-mediated endothelial cell cycle acceleration drives arteriovenous malformations in hereditary hemorrhagic telangiectasia

doi: 10.1101/2023.09.15.554413

Figure Lengend Snippet: ( A ) Heatmap analysis of a high-throughput cell cycle protein ELISA of ECs isolated from livers of P6 BMP9/10ib mice vs. PBS control littermates. Data represent n=2 independent analyses (n represents one litter of pups combined for each condition; PBS, n=6 mice; BMP9/10ib, n=7 mice). ( B ) qPCR analysis of ECs isolated from livers of P6 PBS and BMP9/10ib mice (n=5/condition). Data represent mean ± SEM, Mann-Whitney test ( Cdk2 ), and unpaired t-test ( Cdk4 and Cdk6 ). ( C ) Flow cytometry quantification of p-RB1 fluorescence intensity in ECs isolated from livers of P6 PBS and BMP9/10ib mice (n=3/condition). Data represent mean ± SEM, unpaired t-test. *P < 0.05. ( D and E ) Representative IF staining of p-RB1 (green) and IB4 (red) in the peri-optic nerve and mid-plexus regions ( D ) and corresponding quantification of the peri-optic nerve region ( E ) of retinas from vehicle-treated PBS (n=6), palbociclib-treated PBS (n=5), vehicle-treated BMP9/10ib (n=6), and palbociclib-treated BMP9/10ib (n=6) mice. Data in ( E ) represent individual retinas and mean ± SEM, one-way ANOVA with Tukey’s multiple comparisons test. ns, not significant; *P < 0.05, **P < 0.01. Scale bars in ( D ), 50 μm. ( F ) Representative H&E and IHC staining of p-RB1 in 4 μm skin sections of HHT2 patients. Te, telangiectasia; N, normal vessel; Ba, basilar cells.

Article Snippet: BMP9 (25 mg/kg; MAB3209, R&D System) and BMP10 (50 mg/kg; MAB2926, R&D System) were injected intraperitoneally (i.p.) at P3 and P4.

Techniques: High Throughput Screening Assay, Enzyme-linked Immunosorbent Assay, Isolation, Control, MANN-WHITNEY, Flow Cytometry, Fluorescence, Staining, Immunohistochemistry

Journal: bioRxiv

Article Title: CDK6-mediated endothelial cell cycle acceleration drives arteriovenous malformations in hereditary hemorrhagic telangiectasia

doi: 10.1101/2023.09.15.554413

Figure Lengend Snippet: ( A ) Representative staining using IB4 (green) and of α-smooth muscle actin (SMA, red) in whole petals (first two rows), th vein front (third row), and peri-optic nerve (forth row) and mid-plexus (last two rows) regions of retinas from mice treated as indicated. Arrows denote AVMs; a, artery; v, vein. Scale bars, 1 mm (whole petal images), 100 μm (vein front), and 50 μm (mid-plexus and peri-optic nerve areas). ( B-F ) Scatter plots showing retinal AVM number ( B ), vein diameter ( E ), and mid-plexus vascular density ( F ) following palbociclib treatment, and retinal AVM number ( C ) and AVM diameter ( D ) following ribociclib treatment. Data represent individual retinas and mean ± SEM; vehicle-treated PBS (n=6 mice), palbociclib-treated PBS (n=7), vehicle-treated BMP9/10ib (n=10-12), and palbociclib-treated BMP9/10ib (n=9-13). Data represent individual retinas and mean ± SEM; unpaired t-test ( B-D ), one-way ANOVA with Tukey’s multiple comparisons test ( E and F ). *P 0.05, **P 0.01, ****P 0.0001.

Article Snippet: BMP9 (25 mg/kg; MAB3209, R&D System) and BMP10 (50 mg/kg; MAB2926, R&D System) were injected intraperitoneally (i.p.) at P3 and P4.

Techniques: Staining

Journal: bioRxiv

Article Title: CDK6-mediated endothelial cell cycle acceleration drives arteriovenous malformations in hereditary hemorrhagic telangiectasia

doi: 10.1101/2023.09.15.554413

Figure Lengend Snippet: ( A ) Representative images of tdTomato-stained retinal vasculature from vehicle-treated and palbociclib-treated P6 Eng iECKO mice. Arrows denote AVMs. Scale bars, 1 mm. ( B-D ) Scatter plot showing AVM number ( B ), AVM diameter ( C ), and mid-plexus vascular density ( D ) in retinas of mice treated as in ( A ). Data represent individual retinas and mean ± SEM; vehicle-treated Eng iECKO (n=6 mice), palbociclib-treated Eng iECKO (n=7); unpaired t-test. **P 0.01, ***P 0.001. ( E ) Representative bright field ( a, b, e, f, i, j ) and BABB-cleared ( c, d, g, h, k, l ) images of blue latex bead-perfused P8 PBS control and BMP9/10ib brains, treated or not (vehicle) with palbociclib. First row, dorsal views; second row, ventral views. Arrows in ( d ) denote the two positions where basilar artery (BA) diameter wa measured. ( F ) Quantification of BA diameter across three groups: PBS (n=4), vehicle-treated BMP9/10ib (n=6), and palbociclib-treated BMP9/10ib (n=4) mice. Data represent two measurements per brain and mean ± SEM; one-way ANOVA with Tukey’s multiple-comparisons test. **P 0.01, ***P 0.001.

Article Snippet: BMP9 (25 mg/kg; MAB3209, R&D System) and BMP10 (50 mg/kg; MAB2926, R&D System) were injected intraperitoneally (i.p.) at P3 and P4.

Techniques: Staining, Control

Journal: bioRxiv

Article Title: CDK6-mediated endothelial cell cycle acceleration drives arteriovenous malformations in hereditary hemorrhagic telangiectasia

doi: 10.1101/2023.09.15.554413

Figure Lengend Snippet: ( A ) Representative staining using IB4 (green) and of SMA (red) in whole petals (first two rows), the vein front (third row), and peri-optic nerve (forth row) and mid-plexus (last two rows) regions of retinas from Cdk6 f/f controls and Cdk6 iECKO mice challenged or not (PBS) with BMP9/10ib. Arrows denote AVMs; a, artery; v, vein. Scale bars, 1 mm (whole petal images), 100 μm (vein front), and 50 μm (mid-plexus and peri-optic nerve areas). ( B-D ) Scatter plots showing retinal AVM number ( B ), vein diameter ( C ), and mid-plexus vascular density ( D ) in Cdk6 f/f ;BMP9/10ib controls (n=11-14) and Cdk6 iECKO ;BMP9/10ib (n=8-12) mice. Data represent individual retinas and mean ± SEM, unpaired t-test. **P 0.01, ***P 0.001. ( D ) Schematic illustration of the proposed mechanism of control of the cell cycle in ECs by ALK1 signaling and its relevance for HHT pathogenesis.

Article Snippet: BMP9 (25 mg/kg; MAB3209, R&D System) and BMP10 (50 mg/kg; MAB2926, R&D System) were injected intraperitoneally (i.p.) at P3 and P4.

Techniques: Staining, Control