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R&D Systems wnt5a
Primers used to detect mouse genes

Wnt5a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/wnt5a/product/R&D Systems
Average 94 stars, based on 252 article reviews

wnt5a - by Bioz Stars, 2026-04

94/100 stars

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1) Product Images from "Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury"

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

Journal: Cell Biology and Toxicology

doi: 10.1007/s10565-024-09956-4

Figure Legend Snippet: Primers used to detect mouse genes

Techniques Used:

Desmin-positive cells increase Fzd2/7 while proliferating hepatocytes increase Wnt3a and Wnt5a following acute liver injury. WT C57BL/6 J mice were challenged with CCl 4 or OO and tissues were harvested up to 72 h later. A ) Frozen liver sections were immunostained with Fzd2 or Fzd7 (green) or Desmin (red); images were quantified using Image J and co-localization of Fzd2 or Fzd7 and Desmin was determined by quantifying yellow pixels in Desmin-positive cells. B ) Integrated density of Wnt3a (red) or Wnt5a (red) expression in liver tissues. White boxes denote magnified regions demonstrating co-localized expression. N = 4–5 mice per group. *P < 0.05, **P < 0.01, ***P < 0.001

Figure Legend Snippet: Desmin-positive cells increase Fzd2/7 while proliferating hepatocytes increase Wnt3a and Wnt5a following acute liver injury. WT C57BL/6 J mice were challenged with CCl 4 or OO and tissues were harvested up to 72 h later. A ) Frozen liver sections were immunostained with Fzd2 or Fzd7 (green) or Desmin (red); images were quantified using Image J and co-localization of Fzd2 or Fzd7 and Desmin was determined by quantifying yellow pixels in Desmin-positive cells. B ) Integrated density of Wnt3a (red) or Wnt5a (red) expression in liver tissues. White boxes denote magnified regions demonstrating co-localized expression. N = 4–5 mice per group. *P < 0.05, **P < 0.01, ***P < 0.001

Techniques Used: Expressing

Proinflammatory cytokine mediators are increased following acute liver injury and stimulate Wnt3a and Wnt5a production by hepatocytes and Kupffer cells. WT C57BL/6 J mice were challenged with CCl 4 or OO and tissues were harvested up to 72 h later. A ) Expression of proinflammatory marker mRNA ( tnfα, il-6, and il-10 ) was detected in mHSCs by qRT-PCR. B ) AML12 cells and C ) ImKCs were cultured in the presence or absence of tnf-α or il-6 for 24 h then fixed on glass coverslips and immunostained for Wnt3a and Wnt5a. Immunofluorescence was quantified using ImageJ; relative expression is denoted as arbitrary units of density. Data are from four independent experiments N = 3–4 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Figure Legend Snippet: Proinflammatory cytokine mediators are increased following acute liver injury and stimulate Wnt3a and Wnt5a production by hepatocytes and Kupffer cells. WT C57BL/6 J mice were challenged with CCl 4 or OO and tissues were harvested up to 72 h later. A ) Expression of proinflammatory marker mRNA ( tnfα, il-6, and il-10 ) was detected in mHSCs by qRT-PCR. B ) AML12 cells and C ) ImKCs were cultured in the presence or absence of tnf-α or il-6 for 24 h then fixed on glass coverslips and immunostained for Wnt3a and Wnt5a. Immunofluorescence was quantified using ImageJ; relative expression is denoted as arbitrary units of density. Data are from four independent experiments N = 3–4 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Techniques Used: Expressing, Marker, Quantitative RT-PCR, Cell Culture, Immunofluorescence

Spontaneous activation of mouse hepatic stellate cells is associated with increased expression of ECM genes, Wnt transducers, and β-catenin. A ) Primary mouse hepatic stellate cells were isolated from livers of Wild-type C57BL/6 J mice and cultured for up to 10 days. B ) Expression of activation marker mRNA ( col1a1 and α-sma ) was detected in mHSCs by qRT-PCR. C ) Expression of Wnt transducer mRNA ( fzd1, fzd2, fzd7 , and lrp6) was detected in mHSCs by qRT-PCR. Data are expressed as mHSCs cultured to day 7 and normalized to day 0.5. Data are significantly different from day 0.5, # P < 0.05. D ) mHSCs were fixed on glass coverslips and immunostained for β-catenin and Desmin. Immunofluorescence was quantified using ImageJ; relative expression is denoted as arbitrary units of density. E ) Expression of wnt3a and wnt5a mRNA was determined by qRT-PCR. Data are from three independent experiments, N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Figure Legend Snippet: Spontaneous activation of mouse hepatic stellate cells is associated with increased expression of ECM genes, Wnt transducers, and β-catenin. A ) Primary mouse hepatic stellate cells were isolated from livers of Wild-type C57BL/6 J mice and cultured for up to 10 days. B ) Expression of activation marker mRNA ( col1a1 and α-sma ) was detected in mHSCs by qRT-PCR. C ) Expression of Wnt transducer mRNA ( fzd1, fzd2, fzd7 , and lrp6) was detected in mHSCs by qRT-PCR. Data are expressed as mHSCs cultured to day 7 and normalized to day 0.5. Data are significantly different from day 0.5, # P < 0.05. D ) mHSCs were fixed on glass coverslips and immunostained for β-catenin and Desmin. Immunofluorescence was quantified using ImageJ; relative expression is denoted as arbitrary units of density. E ) Expression of wnt3a and wnt5a mRNA was determined by qRT-PCR. Data are from three independent experiments, N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Techniques Used: Activation Assay, Expressing, Isolation, Cell Culture, Marker, Quantitative RT-PCR, Immunofluorescence

Exogenous Wnt3a and Wnt5a accelerate the spontaneous activation of mouse hepatic stellate cells. A ) Primary mouse hepatic stellate cells were isolated then cultured in the presence or absence of 10 ng/mL of exogenous TGF-β, Wnt3a, and Wnt5a from day 5 to day 7. B ) Expression of col1a1 mRNA was detected in mHSCs by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA and HSC70 (loading control) were measured by Western blot. D ) β-catenin and Desmin expression was analyzed by immunofluorescence in cells following TGF-β, Wnt3a, and Wnt5a treatment. Red arrows highlight nuclear localization of β-catenin. E ) β-catenin expression and nuclear localization, and Desmin expression by immunofluorescence was quantified using ImageJ software. F ) Expression of axin2, ctgf, cyclind1, mmp9, mmp13, and timp1 mRNA and G ) wnt3a and wnt5a mRNA was detected by qRT-PCR. N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001

Figure Legend Snippet: Exogenous Wnt3a and Wnt5a accelerate the spontaneous activation of mouse hepatic stellate cells. A ) Primary mouse hepatic stellate cells were isolated then cultured in the presence or absence of 10 ng/mL of exogenous TGF-β, Wnt3a, and Wnt5a from day 5 to day 7. B ) Expression of col1a1 mRNA was detected in mHSCs by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA and HSC70 (loading control) were measured by Western blot. D ) β-catenin and Desmin expression was analyzed by immunofluorescence in cells following TGF-β, Wnt3a, and Wnt5a treatment. Red arrows highlight nuclear localization of β-catenin. E ) β-catenin expression and nuclear localization, and Desmin expression by immunofluorescence was quantified using ImageJ software. F ) Expression of axin2, ctgf, cyclind1, mmp9, mmp13, and timp1 mRNA and G ) wnt3a and wnt5a mRNA was detected by qRT-PCR. N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001

Techniques Used: Activation Assay, Isolation, Cell Culture, Expressing, Quantitative RT-PCR, SDS Page, Control, Western Blot, Immunofluorescence, Software

HSC activation, via LRP6, occurs in a canonical Wnt-dependent manner. A ) Primary mouse HSCs were then cultured in the presence or absence of TGF-β, Wnt3a, and Wnt5a and the LRP-6 antagonist DKK-1 (10 ng/mL) from day 5 to day 7. B ) Expression of col1a1 mRNA was determined by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA expression and HSC70 (loading control) were measured by Western Blot. α-SMA was normalized to HSC70 as a loading control and quantified using ImageJ software. D ) Expression of axin2, mmp9, and mmp13 mRNA; and E ) wnt3a and wnt5a mRNA was determined in mHSCs by qRT-PCR. Data are from three independent experiments. N = 3–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Figure Legend Snippet: HSC activation, via LRP6, occurs in a canonical Wnt-dependent manner. A ) Primary mouse HSCs were then cultured in the presence or absence of TGF-β, Wnt3a, and Wnt5a and the LRP-6 antagonist DKK-1 (10 ng/mL) from day 5 to day 7. B ) Expression of col1a1 mRNA was determined by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA expression and HSC70 (loading control) were measured by Western Blot. α-SMA was normalized to HSC70 as a loading control and quantified using ImageJ software. D ) Expression of axin2, mmp9, and mmp13 mRNA; and E ) wnt3a and wnt5a mRNA was determined in mHSCs by qRT-PCR. Data are from three independent experiments. N = 3–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Techniques Used: Activation Assay, Cell Culture, Expressing, Quantitative RT-PCR, SDS Page, Control, Western Blot, Software

Mechanism of Wnt-induced HSC reprogramming during wound healing. Acute CCl 4 -induced liver injury damages pericentral hepatocytes and induces innate immune activation in the liver. Proinflammatory mediators, derived from immune cells, including TNF-α and IL-6, perform paracrine-mediated stimulation of resident parenchymal and non-parenchymal cells in the liver. Specifically, hepatocytes and Kupffer cells, in response to inflammatory stimuli, produce Wnt3a and Wnt5a in the microenvironment which in turn, signal to hepatic stellate cells (HSCs). Canonical Wnt-mediated β-catenin activation, signaling via Fzd 1, 2, and 7, drive fibrogenic gene expression leading HSC transdifferentiation to a myofibroblast phenotype to support liver regeneration following acute injury. WNT-; wingless; Fzd- frizzled receptor; LRP6 – low-density lipoprotein receptor-related protein 6; DKK-1 – dickkopf-1; col1a1- collagen 1; α-sma- smooth muscle actin alpha; ECM – extracellular matrix

Figure Legend Snippet: Mechanism of Wnt-induced HSC reprogramming during wound healing. Acute CCl 4 -induced liver injury damages pericentral hepatocytes and induces innate immune activation in the liver. Proinflammatory mediators, derived from immune cells, including TNF-α and IL-6, perform paracrine-mediated stimulation of resident parenchymal and non-parenchymal cells in the liver. Specifically, hepatocytes and Kupffer cells, in response to inflammatory stimuli, produce Wnt3a and Wnt5a in the microenvironment which in turn, signal to hepatic stellate cells (HSCs). Canonical Wnt-mediated β-catenin activation, signaling via Fzd 1, 2, and 7, drive fibrogenic gene expression leading HSC transdifferentiation to a myofibroblast phenotype to support liver regeneration following acute injury. WNT-; wingless; Fzd- frizzled receptor; LRP6 – low-density lipoprotein receptor-related protein 6; DKK-1 – dickkopf-1; col1a1- collagen 1; α-sma- smooth muscle actin alpha; ECM – extracellular matrix

Techniques Used: Activation Assay, Derivative Assay, Expressing

Image Search Results

In vitro experiments showing that PLCB4 in the ProImmuML signature may be a downstream regulator of the Wnt pathway and inhibit GBM proliferation. (A) Metascape analysis of the ProImmuML signature. (B) qPCR analysis was performed to analyze the effects of three different Wnt pathway agonists/inhibitors on the expression of APCDD1 (left panel), RAC2 (middle panel), and PLCB4 (right panel). SKL: SKL-2001, Wnt/β-catenin signaling pathway agonist; MSAB: MSAB, Wnt/β-catenin signaling pathway inhibitor; 2-APB: 2-APB, Wnt/Ca 2+ signaling pathway inhibitor; LON: Lonomycin, Wnt/Ca 2+ signaling pathway agonist; BLE: Blebbistatin, Planar cell polarity pathway inhibitor; WNT: Wnt5a, Planar cell polarity pathway agonist. (C) Representative IHC staining images of PLCB4 from four glioma patients with higher expression of PLCB4 (left panel, n = 2) and lower expression of PLCB4 (right panel, n = 2). (D) The intersection of genes from two parallel comparisons identified 235 upregulated genes (left panel) and 65 downregulated genes (right panel). (E) KEGG enrichment analysis of the upregulated genes. (F) GSEA revealing proliferation-related pathways were significantly inhibited after overexpressing PLCB4. (G) EdU assay showing PLCB4 inhibiting the proliferation of glioma cells in U87. Significant difference, *P<0.05, **P<0.01, ***P<0.001.

Journal: Frontiers in Immunology

Article Title: Meta-analysis of multi-center transcriptomic profiles and machine learning reveal phospholipase Cβ4 as a Wnt/Ca² + signaling mediator in glioblastoma immunotherapy

doi: 10.3389/fimmu.2025.1610683

Figure Lengend Snippet: In vitro experiments showing that PLCB4 in the ProImmuML signature may be a downstream regulator of the Wnt pathway and inhibit GBM proliferation. (A) Metascape analysis of the ProImmuML signature. (B) qPCR analysis was performed to analyze the effects of three different Wnt pathway agonists/inhibitors on the expression of APCDD1 (left panel), RAC2 (middle panel), and PLCB4 (right panel). SKL: SKL-2001, Wnt/β-catenin signaling pathway agonist; MSAB: MSAB, Wnt/β-catenin signaling pathway inhibitor; 2-APB: 2-APB, Wnt/Ca 2+ signaling pathway inhibitor; LON: Lonomycin, Wnt/Ca 2+ signaling pathway agonist; BLE: Blebbistatin, Planar cell polarity pathway inhibitor; WNT: Wnt5a, Planar cell polarity pathway agonist. (C) Representative IHC staining images of PLCB4 from four glioma patients with higher expression of PLCB4 (left panel, n = 2) and lower expression of PLCB4 (right panel, n = 2). (D) The intersection of genes from two parallel comparisons identified 235 upregulated genes (left panel) and 65 downregulated genes (right panel). (E) KEGG enrichment analysis of the upregulated genes. (F) GSEA revealing proliferation-related pathways were significantly inhibited after overexpressing PLCB4. (G) EdU assay showing PLCB4 inhibiting the proliferation of glioma cells in U87. Significant difference, *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Lonomycin (LON; TargetMol, China) and 2-APB (TargetMol, China) served as the activator and inhibitor of the Wnt/Ca2 + pathway, while Wnt5a (TargetMol, China) and blebbistatin (TargetMol, China) were used to modulate the Wnt/PCP pathway.

Techniques: In Vitro, Expressing, Immunohistochemistry, EdU Assay

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Primers used to detect mouse genes

Article Snippet: To measure accelerated HSC activation in vitro, mHSCs were cultured for 7 days and challenged with 10 ng/mL of recombinant mouse TGF –β (R&D Systems #7666-MB) as a positive control of HSC activation, or recombinant mouse Wnt3a (R&D Systems #1324-WN) or Wnt5a (R&D Systems #645-WN), DKK-1 (R&D Systems #5897-DK) on day 5.

Techniques:

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Desmin-positive cells increase Fzd2/7 while proliferating hepatocytes increase Wnt3a and Wnt5a following acute liver injury. WT C57BL/6 J mice were challenged with CCl 4 or OO and tissues were harvested up to 72 h later. A ) Frozen liver sections were immunostained with Fzd2 or Fzd7 (green) or Desmin (red); images were quantified using Image J and co-localization of Fzd2 or Fzd7 and Desmin was determined by quantifying yellow pixels in Desmin-positive cells. B ) Integrated density of Wnt3a (red) or Wnt5a (red) expression in liver tissues. White boxes denote magnified regions demonstrating co-localized expression. N = 4–5 mice per group. *P < 0.05, **P < 0.01, ***P < 0.001

Techniques: Expressing

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Proinflammatory cytokine mediators are increased following acute liver injury and stimulate Wnt3a and Wnt5a production by hepatocytes and Kupffer cells. WT C57BL/6 J mice were challenged with CCl 4 or OO and tissues were harvested up to 72 h later. A ) Expression of proinflammatory marker mRNA ( tnfα, il-6, and il-10 ) was detected in mHSCs by qRT-PCR. B ) AML12 cells and C ) ImKCs were cultured in the presence or absence of tnf-α or il-6 for 24 h then fixed on glass coverslips and immunostained for Wnt3a and Wnt5a. Immunofluorescence was quantified using ImageJ; relative expression is denoted as arbitrary units of density. Data are from four independent experiments N = 3–4 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Techniques: Expressing, Marker, Quantitative RT-PCR, Cell Culture, Immunofluorescence

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Spontaneous activation of mouse hepatic stellate cells is associated with increased expression of ECM genes, Wnt transducers, and β-catenin. A ) Primary mouse hepatic stellate cells were isolated from livers of Wild-type C57BL/6 J mice and cultured for up to 10 days. B ) Expression of activation marker mRNA ( col1a1 and α-sma ) was detected in mHSCs by qRT-PCR. C ) Expression of Wnt transducer mRNA ( fzd1, fzd2, fzd7 , and lrp6) was detected in mHSCs by qRT-PCR. Data are expressed as mHSCs cultured to day 7 and normalized to day 0.5. Data are significantly different from day 0.5, # P < 0.05. D ) mHSCs were fixed on glass coverslips and immunostained for β-catenin and Desmin. Immunofluorescence was quantified using ImageJ; relative expression is denoted as arbitrary units of density. E ) Expression of wnt3a and wnt5a mRNA was determined by qRT-PCR. Data are from three independent experiments, N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Techniques: Activation Assay, Expressing, Isolation, Cell Culture, Marker, Quantitative RT-PCR, Immunofluorescence

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Exogenous Wnt3a and Wnt5a accelerate the spontaneous activation of mouse hepatic stellate cells. A ) Primary mouse hepatic stellate cells were isolated then cultured in the presence or absence of 10 ng/mL of exogenous TGF-β, Wnt3a, and Wnt5a from day 5 to day 7. B ) Expression of col1a1 mRNA was detected in mHSCs by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA and HSC70 (loading control) were measured by Western blot. D ) β-catenin and Desmin expression was analyzed by immunofluorescence in cells following TGF-β, Wnt3a, and Wnt5a treatment. Red arrows highlight nuclear localization of β-catenin. E ) β-catenin expression and nuclear localization, and Desmin expression by immunofluorescence was quantified using ImageJ software. F ) Expression of axin2, ctgf, cyclind1, mmp9, mmp13, and timp1 mRNA and G ) wnt3a and wnt5a mRNA was detected by qRT-PCR. N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001

Techniques: Activation Assay, Isolation, Cell Culture, Expressing, Quantitative RT-PCR, SDS Page, Control, Western Blot, Immunofluorescence, Software

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: HSC activation, via LRP6, occurs in a canonical Wnt-dependent manner. A ) Primary mouse HSCs were then cultured in the presence or absence of TGF-β, Wnt3a, and Wnt5a and the LRP-6 antagonist DKK-1 (10 ng/mL) from day 5 to day 7. B ) Expression of col1a1 mRNA was determined by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA expression and HSC70 (loading control) were measured by Western Blot. α-SMA was normalized to HSC70 as a loading control and quantified using ImageJ software. D ) Expression of axin2, mmp9, and mmp13 mRNA; and E ) wnt3a and wnt5a mRNA was determined in mHSCs by qRT-PCR. Data are from three independent experiments. N = 3–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Techniques: Activation Assay, Cell Culture, Expressing, Quantitative RT-PCR, SDS Page, Control, Western Blot, Software

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Mechanism of Wnt-induced HSC reprogramming during wound healing. Acute CCl 4 -induced liver injury damages pericentral hepatocytes and induces innate immune activation in the liver. Proinflammatory mediators, derived from immune cells, including TNF-α and IL-6, perform paracrine-mediated stimulation of resident parenchymal and non-parenchymal cells in the liver. Specifically, hepatocytes and Kupffer cells, in response to inflammatory stimuli, produce Wnt3a and Wnt5a in the microenvironment which in turn, signal to hepatic stellate cells (HSCs). Canonical Wnt-mediated β-catenin activation, signaling via Fzd 1, 2, and 7, drive fibrogenic gene expression leading HSC transdifferentiation to a myofibroblast phenotype to support liver regeneration following acute injury. WNT-; wingless; Fzd- frizzled receptor; LRP6 – low-density lipoprotein receptor-related protein 6; DKK-1 – dickkopf-1; col1a1- collagen 1; α-sma- smooth muscle actin alpha; ECM – extracellular matrix

Techniques: Activation Assay, Derivative Assay, Expressing

Single-cell RNA sequencing (scRNA-seq) analysis of the dynamically cultured EO and HEO complexes. a. UMAP visualization of 12 samples across four culture conditions: HEOe (HEO + estrogen), HEOp (HEO + estrogen + progesterone + cAMP), EOe (EO + estrogen), EOp (EO + estrogen + progesterone + cAMP). Endometrial epithelial cell clusters were annotated as main Epi (mEpi), ribosome-related Epi (rEpi), proliferative Epi (pEpi), and ciliated Epi (cEpi). Endometrial stromal cell clusters were annotated as main Str (mStr), ribosome-related Str (rStr), proliferative Str (pStr), and a second proliferative Str cluster (pStr2); the endothelial cell clusters included main endothelial cells (mEndo) and ribosome-related endothelial cells (rEndo). b–c. Relative abundance of epithelial (b) and stromal (c) clusters within their respective compartments across groups.]d. Comparison of the percentage of proliferative epithelial cells (pEpi, left panel) and proliferative stromal cells (pStr, right panel) between the dynamically cultured EO and HEO complexes under estrogen culture.e. Validation of enhanced proliferation in HEO complexes by flow cytometry. Significantly higher proportions of EdU + cells in epithelial cells of HEO vs. EO complexes under estrogen culture.f. Ligand-receptor pairs exhibiting significant differences in communication probability between HEOe and EOe complexes. Pathways of biological interest include WNT7A, WNT5A , BMP6 , and LGALS9. Dot size represents the maximum communication probability across compared groups; dot color indicates statistical significance ( P < 0.05). Cluster abbreviations: Epi (endometrial epithelial cells), Str (endometrial stromal cells) and Endo (endothelial cells).g. Circle plot showing the BMP6, LGALS9, WNT7A, WNT5A signaling networks among different cell types in EOe and HEOe complexes.

Journal: Bioactive Materials

Article Title: Microfluidic chip-integrated vascularized endometrial complexes: Mitochondrial function and paracrine crosstalk enhance regenerative potential

doi: 10.1016/j.bioactmat.2025.08.035

Figure Lengend Snippet: Single-cell RNA sequencing (scRNA-seq) analysis of the dynamically cultured EO and HEO complexes. a. UMAP visualization of 12 samples across four culture conditions: HEOe (HEO + estrogen), HEOp (HEO + estrogen + progesterone + cAMP), EOe (EO + estrogen), EOp (EO + estrogen + progesterone + cAMP). Endometrial epithelial cell clusters were annotated as main Epi (mEpi), ribosome-related Epi (rEpi), proliferative Epi (pEpi), and ciliated Epi (cEpi). Endometrial stromal cell clusters were annotated as main Str (mStr), ribosome-related Str (rStr), proliferative Str (pStr), and a second proliferative Str cluster (pStr2); the endothelial cell clusters included main endothelial cells (mEndo) and ribosome-related endothelial cells (rEndo). b–c. Relative abundance of epithelial (b) and stromal (c) clusters within their respective compartments across groups.]d. Comparison of the percentage of proliferative epithelial cells (pEpi, left panel) and proliferative stromal cells (pStr, right panel) between the dynamically cultured EO and HEO complexes under estrogen culture.e. Validation of enhanced proliferation in HEO complexes by flow cytometry. Significantly higher proportions of EdU + cells in epithelial cells of HEO vs. EO complexes under estrogen culture.f. Ligand-receptor pairs exhibiting significant differences in communication probability between HEOe and EOe complexes. Pathways of biological interest include WNT7A, WNT5A , BMP6 , and LGALS9. Dot size represents the maximum communication probability across compared groups; dot color indicates statistical significance ( P < 0.05). Cluster abbreviations: Epi (endometrial epithelial cells), Str (endometrial stromal cells) and Endo (endothelial cells).g. Circle plot showing the BMP6, LGALS9, WNT7A, WNT5A signaling networks among different cell types in EOe and HEOe complexes.

Article Snippet: The spheroids were stimulated with 100 μL of ECM with different treatments, including WNT5A (200 ng/mL, CSB-EP026138HU, CUSABIO, Wuhan, China), WNT7A (100 ng/mL, P06680 , Solarbio), DKK1 (200 ng/mL, C12B, Novoprotein), BOX5 (HY-123071A, MCE), anti-WNT5A neutralizing antibody (2 μg/mL, MAB645, R&D), anti-WNT7A neutralizing antibody (10 μg/mL, sc-365665, Santa Cruz), or VEGFA (10 ng/mL, C744, Novoprotein) as a positive control, by adding dropwise onto the collagen matrix.

Techniques: RNA Sequencing, Cell Culture, Comparison, Biomarker Discovery, Flow Cytometry

Endometrial epithelial and stromal cells enhance the formation of vascular networks in HUVECs within the HEO complex through WNT7A and WNT5A signaling pathways. a. Representative images of sprouting assays using HUVEC spheroids treated as indicated. HUVEC spheroids cultured with basic ECM medium are shown as the blank group (Control). VEGFA added to the basic ECM medium served as the positive control. Scale bar: 400 μm. b. Quantification of sprouting number was determined by Image J software. ∗ P < 0.05 versus Control; # P < 0.05 versus WNT7A; $ P < 0.05 versus DKK1; ^ P < 0.05 versus WNT5A; & P < 0.05 versus BOX5; one-way ANOVA was conducted for statistical analysis. c. Representative images of the tube formation of HUVECs on Matrigel surface treated as indicated. HUVECs cultured with basic ECM medium are shown as the blank (Control). VEGFA added to the basic ECM medium served as the positive control. Scale bar: 400 μm. d. Quantification of branch number using Image J software. ∗ P < 0.05 versus Control; $ P < 0.05 versus DKK1; &P < 0.05 versus BOX5; one-way ANOVA was conducted for statistical analysis.e. Vascular networks of complex with HUVECs and ESCs (HE), complex with HUVECs and EEOs (HO), and HEO treated with WNT7A (or its inhibitor DKK1) and WNT5A (or its inhibitor BOX5) as indicated. Vascular networks are depicted in green. Scale bar: 100 μm.f. Quantification of the vessel-positive area within the formed vascular network across different groups. ∗P < 0.05, ∗∗P < 0.01, Student's t-test.g. The electrical resistance of HEO and HE with DKK1 and WNT7A treatments, respectively. ∗∗∗P < 0.001, two-way ANOVA.h. The electrical resistance of HEO and HO with BOX5 and WNT5A treatments, respectively. ∗∗∗P < 0.001, two-way ANOVA.

Journal: Bioactive Materials

Article Title: Microfluidic chip-integrated vascularized endometrial complexes: Mitochondrial function and paracrine crosstalk enhance regenerative potential

doi: 10.1016/j.bioactmat.2025.08.035

Figure Lengend Snippet: Endometrial epithelial and stromal cells enhance the formation of vascular networks in HUVECs within the HEO complex through WNT7A and WNT5A signaling pathways. a. Representative images of sprouting assays using HUVEC spheroids treated as indicated. HUVEC spheroids cultured with basic ECM medium are shown as the blank group (Control). VEGFA added to the basic ECM medium served as the positive control. Scale bar: 400 μm. b. Quantification of sprouting number was determined by Image J software. ∗ P < 0.05 versus Control; # P < 0.05 versus WNT7A; $ P < 0.05 versus DKK1; ^ P < 0.05 versus WNT5A; & P < 0.05 versus BOX5; one-way ANOVA was conducted for statistical analysis. c. Representative images of the tube formation of HUVECs on Matrigel surface treated as indicated. HUVECs cultured with basic ECM medium are shown as the blank (Control). VEGFA added to the basic ECM medium served as the positive control. Scale bar: 400 μm. d. Quantification of branch number using Image J software. ∗ P < 0.05 versus Control; $ P < 0.05 versus DKK1; &P < 0.05 versus BOX5; one-way ANOVA was conducted for statistical analysis.e. Vascular networks of complex with HUVECs and ESCs (HE), complex with HUVECs and EEOs (HO), and HEO treated with WNT7A (or its inhibitor DKK1) and WNT5A (or its inhibitor BOX5) as indicated. Vascular networks are depicted in green. Scale bar: 100 μm.f. Quantification of the vessel-positive area within the formed vascular network across different groups. ∗P < 0.05, ∗∗P < 0.01, Student's t-test.g. The electrical resistance of HEO and HE with DKK1 and WNT7A treatments, respectively. ∗∗∗P < 0.001, two-way ANOVA.h. The electrical resistance of HEO and HO with BOX5 and WNT5A treatments, respectively. ∗∗∗P < 0.001, two-way ANOVA.

Techniques: Protein-Protein interactions, Cell Culture, Control, Positive Control, Software

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

Review

Structured Review

Images

1) Product Images from "Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury"

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