Journal: Biomaterials advances

Article Title: Enhancing BMP-2-mediated osteogenesis with a synthetic heparan sulfate mimetic.

doi: 10.1016/j.bioadv.2023.213671

Figure Lengend Snippet: Fig. 2. Glycomimetic 15 binds BMP-2 and several other pro-regenerative factors with high affinity. A – SPR solution competition dosing experiments were performed with Ai - BMP-2, Aii - BMP-4, Aiii - BMP-7, Aiv BMPRII Fc chimera and Av - VEGF165 using a dose range of glycomimetic 15 (3.91 to 8000 nM). Avi – normalized response data extrapolated from sensorgrams Ai-Av. B – fluorescence polarization solution competition with BMP-2 (50 nM), 488-labelled heparin dp24 (0.5 nM) and increasing concentrations of glycomimetic 15 (red) or glycomimetic 18 (blue). Ci – DSF melt curves. Cii – first derivative of melt curve data. Ciii – BMP-2 melt temperature when incubated with multiple concentrations of glycomimetic 15, derived from the first derivative of melt curve data. Di – Normalized anti-factor Xa activity for glycomimetic 15 (black), heparin dp12 (red) and heparin dp24 (blue). Dii – anti-factor Xa activity in IU/mg, calculated using USP heparin standard curve. Bars represent mean, and error bars represent S.D. All data is representative of three independent experiments.

Article Snippet: BMP-2 (cat. # 355-BM/ CF), BMP-4 (cat. # 314-BP/CF), BMP-7 (cat. # 354-BP/CF), BMPRII Fc chimera (cat. # 811-BR), Noggin (cat. # 6057-NG/CF), VEGF165 (cat. # 293-VE/CF) polyclonal Goat anti-Human BMP-2/BMP-4 IgG (cat. # AF355), biotinylated monoclonal Mouse anti-Human BMP-2 IgG (cat. #BAM3552) were purchased from R&D systems.

Techniques: Fluorescence, Incubation, Derivative Assay, Activity Assay

Journal: Biomaterials advances

Article Title: Enhancing BMP-2-mediated osteogenesis with a synthetic heparan sulfate mimetic.

doi: 10.1016/j.bioadv.2023.213671

Figure Lengend Snippet: Fig. 5. Glycomimetic 15 influences BMPRII: BMP-2 and BMP-2: cell surface interactions. Ai – Co-immunoprecipitation and Western blot of BMP-2 (125 nM) with BMPRII Fc chimera (250 nM) ± increasing concentrations of either glycomimetic 15 or 18. Aii – Log2 fold change of densitometry values obtained from three in dependent co-immunoprecipitation experiments – bars represent mean log2 fold change ±95 % CI, normalized to BMPRII + BMP-2 alone. Bi – Flow cytometry histograms of C2C12 cells stained with BMP-2 (black histogram) ± increasing concentrations of glycomimetic 15 (red histogram) or glycomimetic 18 (blue histo gram). Bii – Cell surface binding of BMP-2 to C2C12 cells incubated with increasing concentrations of glycomimetic 15 (red) or glycomimetic 18 (blue) – data is mean geometric mean fluorescence ± S.D. normalized to BMP-2 alone. All data either constitutes (Aii, Bii) or is representative of (Ai, Bi) three independent experiments. Statistical analysis was performed via one-way ANOVA with Tukey’s post hoc testing (Aii) or two-way ANOVA with ˇSíd´ak or Bonferroni post hoc testing (Bii). Sig nificance is denoted by *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.

Article Snippet: BMP-2 (cat. # 355-BM/ CF), BMP-4 (cat. # 314-BP/CF), BMP-7 (cat. # 354-BP/CF), BMPRII Fc chimera (cat. # 811-BR), Noggin (cat. # 6057-NG/CF), VEGF165 (cat. # 293-VE/CF) polyclonal Goat anti-Human BMP-2/BMP-4 IgG (cat. # AF355), biotinylated monoclonal Mouse anti-Human BMP-2 IgG (cat. #BAM3552) were purchased from R&D systems.

Techniques: Immunoprecipitation, Western Blot, Flow Cytometry, Staining, Binding Assay, Incubation, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: Characterization of erythroferrone structural domains relevant to its iron-regulatory function

doi: 10.1016/j.jbc.2023.105374

Figure Lengend Snippet: ERFE and BMP receptors bind BMPs in a similar manner. A , crystallography-resolved structure of BMP2 homodimer bound to ALK3 (PDB: 2QJ9 ). Callout box enlarges the interaction and identifies the contacting residues. B , AlphaFold2 model of a BMP2 homodimer ( tan color) bound to ERFE colored by segment ( left ) and confidence pLDDT ( right ). Callout box shows the analogous interaction that BMPs have with their receptors. C , surface plasmon resonance sensorgrams of competition between WT ERFE, W82A ERFE, and activin RIIA for BMP2 ( left ) and BMP6 ( right ) binding. Extracellular portions of activin RIIA were immobilized, and BMP analyte flowed over alone or mixed with WT or W82A ERFE N-terminal segments. D , surface plasmon resonance of the ERFE N terminus binding to extracellular portions of five BMP receptors. BMP, bone morphogenetic protein; ERFE, erythroferrone; pLDDT, predicted local distance difference test.

Article Snippet: For the SPR competition assay, Activin RIIA-Fc (#340-RC2-100), BMPRIA/ALK3-Fc (#2406-BR-100), BMPRII-Fc (#811-BR-100), Activin RIA/ALK2-Fc (#637-AR-100), Activin RIIB-Fc (#339-RB-100/CF), human IgG1-Fc for control (#110-HG-100), BMP2/6 (#7145-BP-010/CF), BMP2 (#355-BM-010/CF), and BMP6 (507-BP-020/CF) were all obtained from RND Systems.

Techniques: SPR Assay, Binding Assay

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 1. Specificity of anti-BMPRIA, anti-BMPRIB and anti-BMPRII antibodies. (A) Wells containing anti-BMPRIA antibody (IA-ab) show intense ELISA reaction only when coated by BMPRIA protein (IA-ag), but not BMPRIB protein (IB-ag). Wells containing anti-BMPRIB antibody (IB-ab) show intense ELISA reaction only when coated by BMPRIB protein (IB-ag), but not BMPRIA protein (IA-ag). Wells containing anti-BMPRII antibody show intense ELISA reaction only when coated by BMPRII protein (). (B) anti-BMPRIA, anti-BMPRIB and anti-BMPRII antibodies specifically recognize the corresponding proteins in a dose-dependent manner. (C–H) Photomicrographs of pre-absorption tests; control (C–E), pre-absorbed (F–H). Note that immunoreactivities in the layer V of the cortex are completely abolished by pre-absorption tests. Scale bar100 m for (C–H).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 2. BMPRIA, BMPRIB and BMPRII expressions in the rat whole brain. 12, hypoglossal nucleus; Amy, amygdala; AP, area postrema; cc, corpus callosum; Cer, cerebellum; CN, cerebellar nuclei; Cor, cerebral cortex; CPu, caudate putamen; DH, dorsal horn; Hi, hippocampus; IO, inferior olive; Pir, piriform cortex; VC, ventral cochlear nucleus, VH, ventral horn. Scale bar3 mm for (A–I); 2 mm for (J–O).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 3. BMPRIA, BMPRIB and BMPRII expressions in the olfactory bulb (A–I) and medial septal nucleus (J–L). EPl, external plexiform layer; Gl, glomerular layer; Gr, granular layer; MS, medial septal nucleus; ON, olfactory nerve layer; SE, subependymal layer. Scale bar 200 m for (A, D, G); 100 m for (B, C, E, F, H–L).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 4. BMPRIA, BMPRIB and BMPRII expressions in the cerebral cortex (A–D, F–I, K–N) and piriform cortex (E, J, O). I–VI, layers I–VI; Pir, piriform cortex. Scale bar200 m for (A, E, F, J, K, O); 100 m for (B–D, G–I, L–N).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 5. BMPRIA, BMPRIB and BMPRII expressions in the hippocampus. CA1-3, fields CA1-3 of Ammon’s horn; DG, dentate gyrus; Gr, granular layer; LM, stratum lacnosum-moleculare; Mo, stratum moleculare; Or, stratum oriens; Po, polymorphological layer; Py, stratum pyramidale; Ra, stratum radiatum. Scale bar 500 m for (A, D, G); 100 m for (B, E, H, J–L); 50 m for (C, F, I, M–O).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 6. BMPRIA, BMPRIB and BMPRII expressions in the basolateral amygdaloid nucleus (A–C), caudate putamen (D–F), globus pallidus (G–I) and corpus callosum (J–O). BL, basolateral amygdaloid nucleus; cc, corpus callosum; CPu, caudate putamen; dhc, dorsal hippocampal commissure; GP, globus pallidus. Scale bar100 m for (J–L); 50 m for (A–I, M–O).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 7. BMPRIA, BMPRIB and BMPRII expressions in the mediodorsal thalamic nucleus (A–C), reticular thalamic nucleus (D–F), supraoptic nucleus (G–I), red nucleus (J–L), substantia nigra, reticular part (M–O) and interpeduncular nucleus (P–R). IP, interpeduncular nucleus; MD, mediodorsal thalamic nucleus; RN, red nucleus; Rt, reticular thalamic nucleus; SNR, substantia nigra, reticular part; SO, supraoptic nucleus. Scale bar50 m.

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 8. BMPRIA, BMPRIB and BMPRII expressions in the fascial nerve nucleus (A–C), facial nerve (D–F), hypoglossal nucleus (G–I), ambiguus nucleus (J–L), nucleus of the solitary tract (M–O) and mesencephalic trigeminal nucleus (P–R). 7, facial nucleus; 7n, facial nerve; 12, hypoglossal nucleus; Amb, ambiguus nucleus; Me5, mesencephalic trigeminal nucleus; SOL, nucleus of the solitary tract. Scale bar50 m.

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 9. BMPRIA, BMPRIB and BMPRII expressions in the dorsal cochlear nucleus (A, C, E), ventral cochlear nucleus (B, D, F), lateral vestibular nucleus (G–I), vestibulo-cochlear nerve (J–L), inferior colliculus (M–O), olivary pretectal nucleus (P–R) and superior colliculus (S–U). 8n, vestibulo- cochlear nerve; DC, dorsal cochlear nucleus; IC, inferior colliculus; LVe, lateral vestibular nucleus; opt, optic tract nucleus; SC, superior colliculus; VC, ventral cochlear nucleus. Scale bar50 m.

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 10. BMPRIA, BMPRIB and BMPRII expressions in the locus ceruleus (A–C), pyramidal tract (D–F), inferior olive (G, J, M), cerebellar cortex (H, K, N), and cerebellar nuclei (I, L, O). CN, cerebellar nuclei; Gr, granular layer; IO, inferior olive; LC, locus ceruleus; Mol, molecular layer; Pur, Purkinje cell layer; Py, pyramidal tract. Scale bar50 m.

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 11. BMPRIA, BMPRIB and BMPRII expressions in the spinal cord (A–I), subventircular zone (J–L), choroidal plexus (M–O). Ch, choroidal plexus; DH, dorsal horn; VH, ventral horn; SVZ, subventricular zone; wm, white matter. Scale bar500 m for (A–C); 50 m for (D–O).

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques:

Journal: Neuroscience

Article Title: Bone morphogenetic protein receptor expressions in the adult rat brain.

doi: 10.1016/j.neuroscience.2010.12.027

Figure Lengend Snippet: Fig. 12. Double-staining study showing that BMPRIA, BMPRIB and BMPRII positive cells in the corpus callosum (A, D, G) are also positive for GFAP (B, E, H) in a merged photomicrograph (C, F, I). Scale bar20 m.

Article Snippet: Solutions of 4 g/ml of recombinant human BMPRIA (315-BR100; R & D Systems, Inc., Mineapolis, MN, USA; http://www. rndsystems.com), BMPRIB (505-PR-100; R & D Systems, Inc.) and BMPRII (H00000659-Q01; Abnova Coporation Inc., Taiwan; http:// www.abnova.com) in 0.1 M carbonate buffer (pH 9.6) were prepared, 50 l of each antigen solution was added to the corresponding wells (96 Well EIA/RIA Plate; Corning, Inc., Corning, NY, USA), and then incubated for overnight at 4 °C.

Techniques: Double Staining