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soluble ephrinb2-fc fusion protein (b2-fc), containing ectodomain residues 27 to 227 of mouse ephrinb2 (R&D Systems)

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R&D Systems soluble ephrinb2-fc fusion protein (b2-fc), containing ectodomain residues 27 to 227 of mouse ephrinb2
Soluble Ephrinb2 Fc Fusion Protein (B2 Fc), Containing Ectodomain Residues 27 To 227 Of Mouse Ephrinb2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/soluble+ephrinb2+fc/ephrinb2+fc/10__1128_slash_jvi__00076___12-73-13-18
Average 90 stars, based on 1 article reviews

soluble ephrinb2-fc fusion protein (b2-fc), containing ectodomain residues 27 to 227 of mouse ephrinb2 - by Bioz Stars, 2026-09

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Article Title: Pyrazolo[1,5-a]pyridine-3-carboxylic acids as EphB and VEGFR2 kinase inhibitors
Article Snippet: Ligand induced autophosphorylation is induced by the addition of 1 microg/ml soluble ephrinB2-Fc (s-eph-rinB2-Fc: R&D Biosystems, CatNr 496-EB) and 0.1 microM ortho-vanadate.

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LIPUS improves compensatory bone formation in OTM via EphrinB2/EphB4 signaling. (A) Representative images of alveolar bone cortex on the pressure side labelled with Calcein and alizarin red S (ARS) (scale bar = 400 μm). (B) Quantification of mineral apposition rate (MAR) of the control and LIPUS sides, n = 4. (C) Representative immunohistochemical staining images of EphB4 and EphrinB2 at ×200 (left) and ×400 (right) magnification (scale bar = 200 µm). (D) The quantitative analysis of EphB4 and EphrinB2 expression by the positive area and average optical density of the alveolar bone, n = 3. Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001 represent the statistical difference between the indicated groups.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

doi: 10.3389/fbioe.2023.1192720

Figure Lengend Snippet: LIPUS improves compensatory bone formation in OTM via EphrinB2/EphB4 signaling. (A) Representative images of alveolar bone cortex on the pressure side labelled with Calcein and alizarin red S (ARS) (scale bar = 400 μm). (B) Quantification of mineral apposition rate (MAR) of the control and LIPUS sides, n = 4. (C) Representative immunohistochemical staining images of EphB4 and EphrinB2 at ×200 (left) and ×400 (right) magnification (scale bar = 200 µm). (D) The quantitative analysis of EphB4 and EphrinB2 expression by the positive area and average optical density of the alveolar bone, n = 3. Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001 represent the statistical difference between the indicated groups.

Article Snippet: In order to stimulate forward signaling, soluble recombinant mouse ephrinB2-Fc chimera (496-EB, R&D, Minneapolis, MN, United States) was preclustered with His-Tag antibody (MAB050, R&D) and used at a final concentration of 1 μg/mL.

Techniques: Control, Immunohistochemical staining, Staining, Expressing

LIPUS promotes the differentiation of bone marrow mesenchymal stem cells (BMSCs)-derived osteoblasts in co-culture system. (A) Schematic of the LIPUS treatment on the BMSC-derived osteoblasts and bone marrow monocytes (BMMs)-derived osteoclasts in the co-culture system. (B) Representative images of ALP and ARS staining of each group on days 7 and 21 after osteogenic induction, respectively. (C) Alkaline Phosphatase (ALP) activity measurement of cell lysates after osteoinduction for 7 days (left) and quantification of mineralized nodules (right). (D) Real time-quantitative PCR (RT-qPCR) analysis of the relative expression of Runx2, Col1a, Bglap, EphB4 and EphrinB2. (E, F) Relative protein levels of EphB4 and EphrinB2 were determined by Western blotting in BMSCs co-cultured with BMMs in the absence or presence of LIPUS. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

doi: 10.3389/fbioe.2023.1192720

Figure Lengend Snippet: LIPUS promotes the differentiation of bone marrow mesenchymal stem cells (BMSCs)-derived osteoblasts in co-culture system. (A) Schematic of the LIPUS treatment on the BMSC-derived osteoblasts and bone marrow monocytes (BMMs)-derived osteoclasts in the co-culture system. (B) Representative images of ALP and ARS staining of each group on days 7 and 21 after osteogenic induction, respectively. (C) Alkaline Phosphatase (ALP) activity measurement of cell lysates after osteoinduction for 7 days (left) and quantification of mineralized nodules (right). (D) Real time-quantitative PCR (RT-qPCR) analysis of the relative expression of Runx2, Col1a, Bglap, EphB4 and EphrinB2. (E, F) Relative protein levels of EphB4 and EphrinB2 were determined by Western blotting in BMSCs co-cultured with BMMs in the absence or presence of LIPUS. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Techniques: Derivative Assay, Co-Culture Assay, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot, Cell Culture

Effect of LIPUS on differentiation of BMM-derived osteoclasts in the co-culture system. (A–C) TRAP staining and F-actin ring staining of BMM-derived osteoclasts at day seven of osteoclastogenesis (scale bar = 400 µm). (B–D) The count of TRAP-positive mononuclear/multinuclear osteoclasts and the number of osteoclasts with F-actin rings were quantified in each well. (E) RT-qPCR analysis of the relative expression of osteoclastogenesis-related genes including c-fos, Nfatc1, MMP9, Ctsk and EphrinB2. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

doi: 10.3389/fbioe.2023.1192720

Figure Lengend Snippet: Effect of LIPUS on differentiation of BMM-derived osteoclasts in the co-culture system. (A–C) TRAP staining and F-actin ring staining of BMM-derived osteoclasts at day seven of osteoclastogenesis (scale bar = 400 µm). (B–D) The count of TRAP-positive mononuclear/multinuclear osteoclasts and the number of osteoclasts with F-actin rings were quantified in each well. (E) RT-qPCR analysis of the relative expression of osteoclastogenesis-related genes including c-fos, Nfatc1, MMP9, Ctsk and EphrinB2. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Techniques: Derivative Assay, Co-Culture Assay, Staining, Quantitative RT-PCR, Expressing

LIPUS enhances osteogenic differentiation in combination with EphrinB2-Fc simulated forward signal. (A) Schematic of LIPUS and EphrinB2-Fc preclustered with Anti-His-tag treatment on BMSC-derived osteoblass. (B) Representative images of ALP and ARS staining of each group on days 7 and 21 after osteogenic induction, respectively. (C) ALP activity measurement of cell lysates after osteoinduction for 7 days (left) and quantification of mineralized nodules (right). (D) RT-qPCR analysis of the relative expression of osteogenesis-related genes including Runx2, Col1a, Bglap and EphB4. (E, F) Relative protein levels of EphB4 were determined by Western blotting in BMSCs incubated with EphrinB2-Fc in the absence or presence of LIPUS. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

doi: 10.3389/fbioe.2023.1192720

Figure Lengend Snippet: LIPUS enhances osteogenic differentiation in combination with EphrinB2-Fc simulated forward signal. (A) Schematic of LIPUS and EphrinB2-Fc preclustered with Anti-His-tag treatment on BMSC-derived osteoblass. (B) Representative images of ALP and ARS staining of each group on days 7 and 21 after osteogenic induction, respectively. (C) ALP activity measurement of cell lysates after osteoinduction for 7 days (left) and quantification of mineralized nodules (right). (D) RT-qPCR analysis of the relative expression of osteogenesis-related genes including Runx2, Col1a, Bglap and EphB4. (E, F) Relative protein levels of EphB4 were determined by Western blotting in BMSCs incubated with EphrinB2-Fc in the absence or presence of LIPUS. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Techniques: Derivative Assay, Staining, Activity Assay, Quantitative RT-PCR, Expressing, Western Blot, Incubation

EhpB4 depletion impairs the osteogenesis induced by LIPUS and EphrinB2-Fc. (A) RT-qPCR analysis of the relative expression of EphB4 in BMSCs transfected with shEphB4 plasmid. (B–C) Relative protein levels of EphB4 were determined by Western blotting. (D) Immunofluorescence staining of EphB4 after transfection with shEphB4 in BMSCs (scale bars = 75 µm). (E) Representative images of ALP and ARS staining of each group on days 7 and 21 after osteogenic induction, respectively. (F) ALP activity measurement of cell lysates after induction for 7 days (left) and quantification of the mineralized nodules (right) in each group. (G) The images depict the migratory ability of different groups as measured by wound healing from 0 h (upper) to 48 h (mid) and Transwell migration assays (lower) in BMSC-derived osteoblasts (scale bar = 400 μm). (H) Quantitative analysis of relative wound closure and migrated cell number was performed using ImageJ software. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

doi: 10.3389/fbioe.2023.1192720

Figure Lengend Snippet: EhpB4 depletion impairs the osteogenesis induced by LIPUS and EphrinB2-Fc. (A) RT-qPCR analysis of the relative expression of EphB4 in BMSCs transfected with shEphB4 plasmid. (B–C) Relative protein levels of EphB4 were determined by Western blotting. (D) Immunofluorescence staining of EphB4 after transfection with shEphB4 in BMSCs (scale bars = 75 µm). (E) Representative images of ALP and ARS staining of each group on days 7 and 21 after osteogenic induction, respectively. (F) ALP activity measurement of cell lysates after induction for 7 days (left) and quantification of the mineralized nodules (right) in each group. (G) The images depict the migratory ability of different groups as measured by wound healing from 0 h (upper) to 48 h (mid) and Transwell migration assays (lower) in BMSC-derived osteoblasts (scale bar = 400 μm). (H) Quantitative analysis of relative wound closure and migrated cell number was performed using ImageJ software. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Western Blot, Immunofluorescence, Staining, Activity Assay, Migration, Derivative Assay, Software

$YAP participates in the signal transduction of EphrinB2/EphB4 in LIPUS-induced osteogenesis through actin filaments. (A) Co-IP experiments showed the interaction between EphB4 and EphrinB2 in the co-culture system as immunoprecipitation with anti-EphB4 antibody to enrich the protein complex. (B) The binding rate of IgG on BMSC-derived osteoblasts, defined as the rate of osteoblasts with attached EphrinB2-Fc among the total osteoblasts, n = 3. (C) Quantification of the proportion of YAP localized in nucleus in BMSC-derived osteoblasts by immunofluorescence using ImageJ, n = 3. (D) Immunofluorescent staining of crosslink antibody (green) to detect EphrinB2-Fc and F-Actin to show cytoskeleton (red). Lower images (scale bar = 50 μm) are high magnification of the white squares in the corresponding upper images (scale bar = 200 μm). (E) YAP nuclear translocation and stress fibres of the cytoskeleton were determined by immunofluorescence (scale bar = 100 µm). Green: YAP staining, Red: F-actin, Blue: nucleus (DAPI), White arrow: filopodia and lamellipodia. (F) Representative images of ALP staining and ARS staining for mineralized nodules on days 7 and 21 after osteogenic induction, respectively. (G, H) The nuclear-cytoplasmic fraction was separated from BMSC-derived osteoblasts treated with LIPUS or EphrinB2-Fc. Immunoblotting assay showed that LIPUS in combination with EphrinB2-Fc treatment significantly increased YAP nuclear localization and slightly decreased phosphor-YAP in cytoplasm, n = 3. (I) Schematic diagram (created with biorender.com ) shows that EphrinB2/EphB4 is involved in regulating migration and osteogenesis of BMSC-derived osteoblasts in a co-culture system under mechanical stimulation with LIPUS. Mechanistically, EphrinB2/EphB4 functions as mechanotransducer in response to the piezoelectric and micro-vibration effect of LIPUS and regulates YAP nuclear translocation through F-actin filament rearrangement. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.$

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

doi: 10.3389/fbioe.2023.1192720

Figure Lengend Snippet: YAP participates in the signal transduction of EphrinB2/EphB4 in LIPUS-induced osteogenesis through actin filaments. (A) Co-IP experiments showed the interaction between EphB4 and EphrinB2 in the co-culture system as immunoprecipitation with anti-EphB4 antibody to enrich the protein complex. (B) The binding rate of IgG on BMSC-derived osteoblasts, defined as the rate of osteoblasts with attached EphrinB2-Fc among the total osteoblasts, n = 3. (C) Quantification of the proportion of YAP localized in nucleus in BMSC-derived osteoblasts by immunofluorescence using ImageJ, n = 3. (D) Immunofluorescent staining of crosslink antibody (green) to detect EphrinB2-Fc and F-Actin to show cytoskeleton (red). Lower images (scale bar = 50 μm) are high magnification of the white squares in the corresponding upper images (scale bar = 200 μm). (E) YAP nuclear translocation and stress fibres of the cytoskeleton were determined by immunofluorescence (scale bar = 100 µm). Green: YAP staining, Red: F-actin, Blue: nucleus (DAPI), White arrow: filopodia and lamellipodia. (F) Representative images of ALP staining and ARS staining for mineralized nodules on days 7 and 21 after osteogenic induction, respectively. (G, H) The nuclear-cytoplasmic fraction was separated from BMSC-derived osteoblasts treated with LIPUS or EphrinB2-Fc. Immunoblotting assay showed that LIPUS in combination with EphrinB2-Fc treatment significantly increased YAP nuclear localization and slightly decreased phosphor-YAP in cytoplasm, n = 3. (I) Schematic diagram (created with biorender.com ) shows that EphrinB2/EphB4 is involved in regulating migration and osteogenesis of BMSC-derived osteoblasts in a co-culture system under mechanical stimulation with LIPUS. Mechanistically, EphrinB2/EphB4 functions as mechanotransducer in response to the piezoelectric and micro-vibration effect of LIPUS and regulates YAP nuclear translocation through F-actin filament rearrangement. Data are presented as the mean ± SEM, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 represent the statistical difference between the indicated groups.

Techniques: Transduction, Co-Immunoprecipitation Assay, Co-Culture Assay, Immunoprecipitation, Binding Assay, Derivative Assay, Immunofluorescence, Staining, Translocation Assay, Western Blot, Migration

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soluble ephrinb2-fc fusion protein (b2-fc), containing ectodomain residues 27 to 227 of mouse ephrinb2 (R&D Systems)

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