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R&D Systems wnt protein
Wnt Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/wnt protein/product/R&D Systems
Average 93 stars, based on 22 article reviews

wnt protein - by Bioz Stars, 2026-03

93/100 stars

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Wnt5a and DPP are present in the ECM of DPSCs and transported via Exosomes. ( A ) Representative confocal micrographs of ECM isolated from DPSCs immunostained for DPP (red) and Wnt5a (green). ( B ) Representative unstained TEM images of exosomes isolated from DPSCs showing the presence of DPP (black arrows; 20 nm gold particles) and Wnt5a (White arrows; 10 nm gold particles).

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: Wnt5a and DPP are present in the ECM of DPSCs and transported via Exosomes. ( A ) Representative confocal micrographs of ECM isolated from DPSCs immunostained for DPP (red) and Wnt5a (green). ( B ) Representative unstained TEM images of exosomes isolated from DPSCs showing the presence of DPP (black arrows; 20 nm gold particles) and Wnt5a (White arrows; 10 nm gold particles).

Article Snippet: In addition, Wnt5a recombinant protein (500ng/ml, R & D Systems or PF-L6 (1nM, a kind gift from AntlerA Therapeutics were used as positive controls to treat DPSCs to monitor nuclear accumulation of β-Catenin.

Techniques: Isolation

DPP stimulation of DPSCs activate Wnt5a signaling. A . DPSCs were treated with DPP (500ng/ml) in the presence and absence of the NF-κB inhibitor TPCA-1. Stimulation was done at varying time points. Total levels of Wnt5a were examined by Western blotting. The densitometric ratios of Wnt5a normalized to actin are shown. B . DPSCs were treated with or without TPCA-1 for 1 h, followed by addition of DPP. The condition media was harvested at the times indicated. WNT5A concentration measured by direct ELISA, and mean and SD ( n = 8) were plotted, *: p < 0.05; ** p < 0.01. C . DPSCs were treated with DMSO (no inhibitor), TPCA-1 and JSH-23. Cells were stimulated with DPP at 500ng/ml for the indicated time points. Expression of Wnt5a gene expression levels were determined by RT-PCR. Fold change was obtained relative to 0 h. D . DPSCs were co-transfected with Wnt5a promoter luciferase plasmids or NF-κB RE luciferase plasmids and treated with DMSO (no inhibitor) and TPCA-1 followed by stimulation with DPP at 0, 250, 500ng/ml. Luciferase activities were measured after 48 h. Mean values with standard deviation for Wnt5a and NF-κB RE promoter activities were normalized to pCMV Renilla and transfection controls to Firefly/Renilla and plotted. Activity was abrogated when cells were treated with TPCA-1. E . ChIP assay was conducted with DPSCs treated with DPP for 0, 1 and 2 h as described in “Materials and Methods” using an anti-NF-κB antibody and the immunoprecipitated DNA was analyzed by qPCR using primers for the NF-κB binding site on the Wnt5a promoter. Mean values with standard deviation for fold enrichments of target DNA fragments, normalized to IgG antibody were plotted. *< 0.05; **<0.01.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: DPP stimulation of DPSCs activate Wnt5a signaling. A . DPSCs were treated with DPP (500ng/ml) in the presence and absence of the NF-κB inhibitor TPCA-1. Stimulation was done at varying time points. Total levels of Wnt5a were examined by Western blotting. The densitometric ratios of Wnt5a normalized to actin are shown. B . DPSCs were treated with or without TPCA-1 for 1 h, followed by addition of DPP. The condition media was harvested at the times indicated. WNT5A concentration measured by direct ELISA, and mean and SD ( n = 8) were plotted, *: p < 0.05; ** p < 0.01. C . DPSCs were treated with DMSO (no inhibitor), TPCA-1 and JSH-23. Cells were stimulated with DPP at 500ng/ml for the indicated time points. Expression of Wnt5a gene expression levels were determined by RT-PCR. Fold change was obtained relative to 0 h. D . DPSCs were co-transfected with Wnt5a promoter luciferase plasmids or NF-κB RE luciferase plasmids and treated with DMSO (no inhibitor) and TPCA-1 followed by stimulation with DPP at 0, 250, 500ng/ml. Luciferase activities were measured after 48 h. Mean values with standard deviation for Wnt5a and NF-κB RE promoter activities were normalized to pCMV Renilla and transfection controls to Firefly/Renilla and plotted. Activity was abrogated when cells were treated with TPCA-1. E . ChIP assay was conducted with DPSCs treated with DPP for 0, 1 and 2 h as described in “Materials and Methods” using an anti-NF-κB antibody and the immunoprecipitated DNA was analyzed by qPCR using primers for the NF-κB binding site on the Wnt5a promoter. Mean values with standard deviation for fold enrichments of target DNA fragments, normalized to IgG antibody were plotted. *< 0.05; **<0.01.

Techniques: Western Blot, Concentration Assay, Direct ELISA, Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Luciferase, Standard Deviation, Activity Assay, Immunoprecipitation, Binding Assay

$DPP stimulation of DPSCs promote nuclear translocation of β-catenin by activating WNT5A. A . Cell fractionation of DPSCs treated with DPP for 0, 0.5, 6 and 24 h were performed. Immunoblotting using anti-βcatenin antibody showed higher amounts of nuclear accumulation as shown by the β-catenin to actin ratio. Tubulin and Lamin A/C were used as controls to demonstrate the purity of cytoplasmic and nuclear extracts. B to E. Immunofluorescence images of DPSC or DPSC/Wnt5a-KO cells treated with DPP for 0, 5, 15, 30 and 60 min. Images were also acquired in the presence of inhibitors TPCA-1& Box5 with DPP stimulation. Note the low levels of nuclear β-catenin in the presence of inhibitors and in Wnt5a-silenced cells. F and G. Immunofluoresence images of DPSCs treated with Wnt5a (positive control) and PF-L6 (positive control for 0,5,15, 30 and 60 min to demonstrate nuclear translocation of β-catenin .DAPI (blue) stains the nucleus. Scale bar 20 μm.$

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: DPP stimulation of DPSCs promote nuclear translocation of β-catenin by activating WNT5A. A . Cell fractionation of DPSCs treated with DPP for 0, 0.5, 6 and 24 h were performed. Immunoblotting using anti-βcatenin antibody showed higher amounts of nuclear accumulation as shown by the β-catenin to actin ratio. Tubulin and Lamin A/C were used as controls to demonstrate the purity of cytoplasmic and nuclear extracts. B to E. Immunofluorescence images of DPSC or DPSC/Wnt5a-KO cells treated with DPP for 0, 5, 15, 30 and 60 min. Images were also acquired in the presence of inhibitors TPCA-1& Box5 with DPP stimulation. Note the low levels of nuclear β-catenin in the presence of inhibitors and in Wnt5a-silenced cells. F and G. Immunofluoresence images of DPSCs treated with Wnt5a (positive control) and PF-L6 (positive control for 0,5,15, 30 and 60 min to demonstrate nuclear translocation of β-catenin .DAPI (blue) stains the nucleus. Scale bar 20 μm.

Techniques: Translocation Assay, Cell Fractionation, Western Blot, Immunofluorescence, Positive Control

DPP activates the expression of odontogenic markers through Wnt5a/β-catenin signaling and abrogates their expression in the presence of Wnt5a/β-catenin pathway inhibitors. DPSCs were cultured under growth conditions and were treated with DMSO; with Box5 or iCRT14. In all conditions cells were stimulated with 500ng/ml DPP and cultured for 0, 2, 6 and 24 h. Total RNA was isolated and quantitative RT-PCR analysis performed. Fold changes were obtained relative to 0 h. Expression levels of early odontogenic markers such as RUNX2 ( A ), OSX ( B ), ALP ( C ), OCN ( D ) and DMP1 (E) increased progressively from 0–24 h. Specificity of Wnt5a/β-catenin signaling was confirmed by gene expression analysis in the presence of Wnt signaling pathway specific inhibitors Box5 and iCRT14. Data are means ± of triplicates. Means and SDs of fold changes to 0 h are shown. Significant difference * p < 0.05; ** p < 0.01.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: DPP activates the expression of odontogenic markers through Wnt5a/β-catenin signaling and abrogates their expression in the presence of Wnt5a/β-catenin pathway inhibitors. DPSCs were cultured under growth conditions and were treated with DMSO; with Box5 or iCRT14. In all conditions cells were stimulated with 500ng/ml DPP and cultured for 0, 2, 6 and 24 h. Total RNA was isolated and quantitative RT-PCR analysis performed. Fold changes were obtained relative to 0 h. Expression levels of early odontogenic markers such as RUNX2 ( A ), OSX ( B ), ALP ( C ), OCN ( D ) and DMP1 (E) increased progressively from 0–24 h. Specificity of Wnt5a/β-catenin signaling was confirmed by gene expression analysis in the presence of Wnt signaling pathway specific inhibitors Box5 and iCRT14. Data are means ± of triplicates. Means and SDs of fold changes to 0 h are shown. Significant difference * p < 0.05; ** p < 0.01.

Techniques: Expressing, Cell Culture, Isolation, Quantitative RT-PCR

DPP stimulation activates receptors and co-receptors of Wnt5a signaling pathway. A . DPSCs were treated with DPP (500ng/ml) for 0, 2, 6, 24 h. Cells were lysed with RIPA buffer and total cell lysates were isolated. Western blotting was performed to detect Wnt5a/β-catenin receptors FZD5, FZD6, ROR2 and co-receptors LRP5 & LRP6. B . DPSCs were treated with varying concentrations of inhibitor Box5 for one hour and stimulated with DPP for 24 h. Western Blotting was performed as above. The density ratios with respect to actin were calculated and indicated on the panel.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: DPP stimulation activates receptors and co-receptors of Wnt5a signaling pathway. A . DPSCs were treated with DPP (500ng/ml) for 0, 2, 6, 24 h. Cells were lysed with RIPA buffer and total cell lysates were isolated. Western blotting was performed to detect Wnt5a/β-catenin receptors FZD5, FZD6, ROR2 and co-receptors LRP5 & LRP6. B . DPSCs were treated with varying concentrations of inhibitor Box5 for one hour and stimulated with DPP for 24 h. Western Blotting was performed as above. The density ratios with respect to actin were calculated and indicated on the panel.

Techniques: Isolation, Western Blot

Spatial and temporal analysis of Wnt5a and its receptors and co-receptors with DPP stimulation using immunofluorescence. A . Visualization of the expression and interaction between Wnt5a (Red, Alexa Fluor 594) and receptor FZD5 (green, Alexa Fluor 488) in DPSCs. Prominent fluorescent spots at 5–30 min indicate Wnt5a-FZD5 interaction in the merged images. Nuclei stained with DAPI. Scale bar:20 μm. B . Visualization of the expression of Wnt5a (Red, Alexa Fluor 594) and receptor FZD5 (green, Alexa Fluor 488) in Wnt5a silenced DPSCs. Note DPP stimulation failed to restore Wnt5a expression and its interaction with FZD5. Scale bar: 20 μm. C . Visualization of the expression and interaction between Wnt5a (Red, Alexa Fluor 594) and co-receptor LRP6 (green, Alexa Fluor 488) in DPSCs. Prominent fluorescent spots at 15–60 min indicate Wnt5a-LRP6 interaction in the merged images. Nuclei stained with DAPI, Scale bar:20 μm. D . Visualization of the expression of Wnt5a (Red, Alexa Fluor 594) and receptor LRP6 (green, Alexa Fluor 488) in Wnt5a silenced DPSCs. Note DPP stimulation failed to restore Wnt5a expression and its interaction with LRP6. Scale bar: 20 μm. E . Visualization of the expression and interaction between Wnt5a (Red, Alexa Fluor 594) and receptor ROR2 (green, Alexa Fluor 488) in DPSCs. Prominent fluorescent spots at 15–60 min indicate Wnt5a-ROR2 interaction in the merged images. Nuclei stained with DAPI. Scale bar:20 μm. F . Visualization of the expression of Wnt5a (Red, Alexa Fluor 594) and receptor FZD5 (green, Alexa Fluor 488) in Wnt5a silenced DPSCs. Note DPP stimulation failed to restore Wnt5a expression and its interaction with ROR2. Scale bar: 20 μm. G Representative TIRF microscopy image showing the presence of Wnt5a (green) & Ror2 (red) on the plasma membrane and their reduced levels with DPP stimulation. Total RNA was isolated from DPSCs stimulated with DPP and quantitative RT-PCR analysis performed. Fold changes were obtained relative to 0 h. Expression levels of Vangl1 ( H ) increased from 0–6 h and Vangl 2 ( I ) progressively from 0–24 h. Means and SDs of fold changes to 0 h are shown. Significant difference ** p < 0.01.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: Spatial and temporal analysis of Wnt5a and its receptors and co-receptors with DPP stimulation using immunofluorescence. A . Visualization of the expression and interaction between Wnt5a (Red, Alexa Fluor 594) and receptor FZD5 (green, Alexa Fluor 488) in DPSCs. Prominent fluorescent spots at 5–30 min indicate Wnt5a-FZD5 interaction in the merged images. Nuclei stained with DAPI. Scale bar:20 μm. B . Visualization of the expression of Wnt5a (Red, Alexa Fluor 594) and receptor FZD5 (green, Alexa Fluor 488) in Wnt5a silenced DPSCs. Note DPP stimulation failed to restore Wnt5a expression and its interaction with FZD5. Scale bar: 20 μm. C . Visualization of the expression and interaction between Wnt5a (Red, Alexa Fluor 594) and co-receptor LRP6 (green, Alexa Fluor 488) in DPSCs. Prominent fluorescent spots at 15–60 min indicate Wnt5a-LRP6 interaction in the merged images. Nuclei stained with DAPI, Scale bar:20 μm. D . Visualization of the expression of Wnt5a (Red, Alexa Fluor 594) and receptor LRP6 (green, Alexa Fluor 488) in Wnt5a silenced DPSCs. Note DPP stimulation failed to restore Wnt5a expression and its interaction with LRP6. Scale bar: 20 μm. E . Visualization of the expression and interaction between Wnt5a (Red, Alexa Fluor 594) and receptor ROR2 (green, Alexa Fluor 488) in DPSCs. Prominent fluorescent spots at 15–60 min indicate Wnt5a-ROR2 interaction in the merged images. Nuclei stained with DAPI. Scale bar:20 μm. F . Visualization of the expression of Wnt5a (Red, Alexa Fluor 594) and receptor FZD5 (green, Alexa Fluor 488) in Wnt5a silenced DPSCs. Note DPP stimulation failed to restore Wnt5a expression and its interaction with ROR2. Scale bar: 20 μm. G Representative TIRF microscopy image showing the presence of Wnt5a (green) & Ror2 (red) on the plasma membrane and their reduced levels with DPP stimulation. Total RNA was isolated from DPSCs stimulated with DPP and quantitative RT-PCR analysis performed. Fold changes were obtained relative to 0 h. Expression levels of Vangl1 ( H ) increased from 0–6 h and Vangl 2 ( I ) progressively from 0–24 h. Means and SDs of fold changes to 0 h are shown. Significant difference ** p < 0.01.

Techniques: Immunofluorescence, Expressing, Staining, Microscopy, Membrane, Isolation, Quantitative RT-PCR

Effect of DPP-mediated Wnt/βcatenin signaling on the terminal differentiation of DPSCs into odontoblastic lineage. DPSCs and DPSC/Wnt5a-KO cells were stimulated with or without DPP and cultured under differentiation conditions for 0–3 weeks. Total RNA was isolated at the indicated time points and quantitative RT-PCR analysis performed. Fold changes were obtained relative to day 0. Expression levels of odontogenic markers such as ( A ) RUNX2 ; ( B ) OSX ; ( C ) ALP ; ( D ) COL1A1 , ( E ) DMP1 , ( F ) FN1 ; ( G ) OCN ; ( H ) OPG ; ( I ) OPN ; ( J ) VEGFA . Gene expression fold changes calculated at the relative ratios to day 0 and means and SDs of changes and comparisons are shown. Note lower gene expression levels in Wnt5a silenced DPSCs even with DPP stimulation. * p < 0.05; ** p < 0.01.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: Effect of DPP-mediated Wnt/βcatenin signaling on the terminal differentiation of DPSCs into odontoblastic lineage. DPSCs and DPSC/Wnt5a-KO cells were stimulated with or without DPP and cultured under differentiation conditions for 0–3 weeks. Total RNA was isolated at the indicated time points and quantitative RT-PCR analysis performed. Fold changes were obtained relative to day 0. Expression levels of odontogenic markers such as ( A ) RUNX2 ; ( B ) OSX ; ( C ) ALP ; ( D ) COL1A1 , ( E ) DMP1 , ( F ) FN1 ; ( G ) OCN ; ( H ) OPG ; ( I ) OPN ; ( J ) VEGFA . Gene expression fold changes calculated at the relative ratios to day 0 and means and SDs of changes and comparisons are shown. Note lower gene expression levels in Wnt5a silenced DPSCs even with DPP stimulation. * p < 0.05; ** p < 0.01.

Techniques: Cell Culture, Isolation, Quantitative RT-PCR, Expressing

Silencing Wnt5a in DPSCs impair their ability to assemble a calcified extracellular matrix. A . DPSCs and DPSC/Wnt5a-KO cells were stimulated with or without 500ng DPP and cultured under differentiation conditions for 0–3 weeks. The mineralized nodules containing calcium were visualized using Alizarin Red staining. Note the small size of the mineralized nodule in the Wnt5a-silenced DPSCs. Insets were scanned images of a well from 12 well tissue culture plate. B . Quantitative measurement of the calcium content in the deposited nodule was determined by measuring the absorbance of the eluted Alizarin Red stain at 562 nm on a multiplate reader using a standard calcium curve. Statistically significant differences are indicated at 1, 2 and 3 weeks.*p, 0.05, **p, 0.01.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: Silencing Wnt5a in DPSCs impair their ability to assemble a calcified extracellular matrix. A . DPSCs and DPSC/Wnt5a-KO cells were stimulated with or without 500ng DPP and cultured under differentiation conditions for 0–3 weeks. The mineralized nodules containing calcium were visualized using Alizarin Red staining. Note the small size of the mineralized nodule in the Wnt5a-silenced DPSCs. Insets were scanned images of a well from 12 well tissue culture plate. B . Quantitative measurement of the calcium content in the deposited nodule was determined by measuring the absorbance of the eluted Alizarin Red stain at 562 nm on a multiplate reader using a standard calcium curve. Statistically significant differences are indicated at 1, 2 and 3 weeks.*p, 0.05, **p, 0.01.

Techniques: Cell Culture, Staining

Expression of Wnt5a and signaling components of Wnt5a in WT & DSPP-KO mice. Post-natal day 3 DSPP null mice and their matched wild type mice heads were used for immunohistochemical analysis. Sections were treated with primary antibodies against β-catenin ( A ); Wnt5a ( B ); Fzd 5( C ); Fzd6( D ); Lrp5 ( E ); Lrp6 ( F ) and Ror2 ( G ). Boxes marked are higher images showing staining in the dental pulp cells.

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: Expression of Wnt5a and signaling components of Wnt5a in WT & DSPP-KO mice. Post-natal day 3 DSPP null mice and their matched wild type mice heads were used for immunohistochemical analysis. Sections were treated with primary antibodies against β-catenin ( A ); Wnt5a ( B ); Fzd 5( C ); Fzd6( D ); Lrp5 ( E ); Lrp6 ( F ) and Ror2 ( G ). Boxes marked are higher images showing staining in the dental pulp cells.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

doi: 10.1038/s41598-024-76069-7

Figure Lengend Snippet: DNA oligoes for quantitative PCR.

Techniques: Sequencing

Journal: iScience

Article Title: Atypical KCNQ1/Kv7 channel function in a neonatal diabetes patient: Hypersecretion preceded the failure of pancreatic β-cells

doi: 10.1016/j.isci.2024.110291

Figure Lengend Snippet:

Article Snippet: Day 0 media: S1/S2 basal media, 100 ng/mL Activin A (R&D Systems), 25 ng/mL mouse Wnt3a (R&D Systems).

Techniques: Control, Purification, Virus, Recombinant, PCR Cloning, Bicinchoninic Acid Protein Assay, Plasmid Preparation, Gel Extraction, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Transformation Assay, Software

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