Journal: bioRxiv

Article Title: Wnt5a-mediated Adipo-Cardiac Interorgan Communication in HFpEF

doi: 10.1101/2025.10.29.685456

Figure Lengend Snippet: (A) Schematic outline of study. (B) Cellular localization of differentially expressed genes (DEGs) in Nprc -AKO visceral adipose tissue (VAT). (C) Increased and decreased gene ontologies in Nprc -AKO vs control VAT. (D) Volcano plot of DEGs in Nprc -AKO vs control VAT (red indicates p < 0.01 and 50%-fold-change compared to control). (E) Reverse transcriptase quantitative PCR of top 5 Wnt isoforms in Nprc -AKO vs control VAT. (F) Plasma circulating levels of Wnt5a in global (GKO), cardiomyocyte-specific (CKO), adipocyte-specific (AKO), and control (WT) mice after L-NAME and high fat diet. (G) Cell size in H9C2 cells after treatment with Wnt5a. (H) RT-qPCR for hypertrophic markers NPPA, NPPB, and MYH7 in H9C2 cells after Wnt5a treatment.

Article Snippet: For studies investigating the effects of Wnt5a treatment, cells were transfected with either pcDNA3.2-Wnt5a plasmid (Addgene 43813) or empty vector control (Addgene 29496).

Techniques: Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Clinical Proteomics, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Wnt5a-mediated Adipo-Cardiac Interorgan Communication in HFpEF

doi: 10.1101/2025.10.29.685456

Figure Lengend Snippet: (A) Schematic outline of study. (B) Histologic and wheat germ agglutinin-based hypertrophy, (C) plasma Wnt5a levels, (D) left ventricular systolic pressure, (E) left ventricular end-diastolic pressure, (F) diastolic relaxation of the left ventricle, (G) right ventricular systolic pressure, (H) right ventricular end-diastolic pressure, (I) diastolic relaxation of the right ventricle, (J) left atrial end-systolic volume by echocardiography, and (K) E/e’ ratio by echocardiography in C57/BL6J mice treated with 8 weeks of L-NAME and high fat diet, followed by LGK974 or carrier control intraperitoneal injections.

Article Snippet: For studies investigating the effects of Wnt5a treatment, cells were transfected with either pcDNA3.2-Wnt5a plasmid (Addgene 43813) or empty vector control (Addgene 29496).

Techniques: Clinical Proteomics, Control

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

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.

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

Journal: Oral oncology

Article Title: Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition.

doi: 10.1016/j.oraloncology.2017.03.019

Figure Lengend Snippet: Fig. 1. Expression of DNp63, Wnt5a, Ror1, and Ror2 in tongue SCC cell lines. (A) RT- PCR analysis shows that DNp63 is not expressed in high metastatic potential SQUU- B cells, but is expressed in other cell lines. Contrastingly, Wnt5a and Ror2 are strongly expressed in SQUU-B cells, but not the other cell lines. Ror1 is equally expressed in all cell lines. (B, C) Real-time PCR and western blotting analyses. DNp63 is expressed in SQUU-BO, but not in SQUU-BC cells. Wnt5a and Ror2 expression in SQUU-BO cells is lower than that in SQUU-BC cells (Mann–Whitney U test, p < 0.05). No significant difference in Ror1 expression between SQUU-BC and SQUU-BO cells is found. (D, E) Real-time PCR and western blotting analyses. DNp63 expression is low in SQUU-A cells transfected with DNp63 siRNA. Wnt5a and Ror2 expression is up-regulated by DNp63 knockdown (Mann–Whitney U test, p < 0.05). However, Ror1 expression is unchanged.

Article Snippet: Exogenous Wnt5a treatment Recombinant human Wnt5a protein (rhWnt5a) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, MANN-WHITNEY, Transfection, Knockdown

Journal: Oral oncology

Article Title: Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition.

doi: 10.1016/j.oraloncology.2017.03.019

Figure Lengend Snippet: Fig. 2. Effects of Wnt5a knockdown on expression of EMT-associated molecules and capacities for migration and invasion. (A) RT-PCR and western blotting analyses demonstrate Wnt5a inhibition following transfection of Wnt5a siRNA into SQUU-B cells. (B) Real-time PCR analyses demonstrate downregulation of CK19 and upregulation of vimentin (Mann–Whitney U test, p < 0.05). (C) Wound healing assay demonstrates low migration capacity 48 h after transfection of Wnt5a siRNA (Mann–Whitney U test, p < 0.05). Scale bar, 200 lm. (D) Invasion assay shows that Wnt5a-knockdown cells have low invasion capacity at 22 h after transfection of Wnt5a siRNA (Mann–Whitney U test, p < 0.05). Scale bars, 100 lm.

Article Snippet: Exogenous Wnt5a treatment Recombinant human Wnt5a protein (rhWnt5a) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Knockdown, Expressing, Migration, Reverse Transcription Polymerase Chain Reaction, Western Blot, Inhibition, Transfection, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Wound Healing Assay, Invasion Assay

Journal: Oral oncology

Article Title: Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition.

doi: 10.1016/j.oraloncology.2017.03.019

Figure Lengend Snippet: Fig. 3. Effects of Ror2 knockdown on expression of EMT-associated molecules and capacities for migration and invasion. (a) RT-PCR and western blotting analyses show inhibition of Ror2 expression following transfection of Ror2 siRNA into SQUU-B cells. (b) Real-time PCR analyses demonstrate downregulation of CK19 and upregulation of Wnt5a (Mann–Whitney U test, p < 0.05). (c) Wound healing assay shows that Ror2-depleted cells have low migration ability at 48 h after transfection of siRNA (Mann– Whitney U test, p < 0.05). Scale bar, 200 lm. (d) Invasion assay shows that Ror2-depleted cells have low invasion ability at 22 h after transfection of siRNA (Mann–Whitney U test, p < 0.05). Scale bars, 100 lm.

Article Snippet: Exogenous Wnt5a treatment Recombinant human Wnt5a protein (rhWnt5a) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Knockdown, Expressing, Migration, Reverse Transcription Polymerase Chain Reaction, Western Blot, Inhibition, Transfection, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Wound Healing Assay, Invasion Assay

Journal: Oral oncology

Article Title: Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition.

doi: 10.1016/j.oraloncology.2017.03.019

Figure Lengend Snippet: Fig. 4. Immunohistochemical detection and cause-specific survival curves in tongue SCC patients. Anti-Wnt5a and -Ror2 antibodies were used to stain normal tongue epithelial tissues and tongue SCC specimens. In the normal epithelial tissues, Wnt5a and Ror2 are almost undetectable (Aa, b). In tongue SCC tissues, Wnt5a and Ror2 are frequently strongly present in the cytoplasm and at the membrane of cancer cells (Ac, d), but are undetectable in some tongue SCC tissues (Ae, f). Scale bars, 50 lm. Patients with high Wnt5a or Ror2 expression have lower cause-specific 5-year survival rates than each low expression group (log-rank test) (B).

Article Snippet: Exogenous Wnt5a treatment Recombinant human Wnt5a protein (rhWnt5a) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Immunohistochemical staining, Staining, Membrane, Expressing

Journal: Oral oncology

Article Title: Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition.

doi: 10.1016/j.oraloncology.2017.03.019

Figure Lengend Snippet: Fig. 5. Expression and gelatinolytic activity of MMP-2 in tongue SCC cells. (A) RT-PCR analyses demonstrate that MMP-2 is more strongly expressed in SQUU-B cells expressing Wnt5a and Ror2 compared with other OSCC cells. (B) Real-time PCR analyses show downregulation of MMP-2 expression following DNp63b overexpression, Wnt5a knockdown, or Ror2 knockdown, and upregulation by DNp63 knockdown (Mann–Whitney U test, p < 0.05). (C) Gelatin zymography assay with upper bands indicating preformed MMP-2 protein and lower bands indicating active MMP-2 protein. Active MMP-2 expression is present in SQUU-B cells but not SQUU-A cells. Expression of preformed and active MMP-2 is increased by DNp63 knockdown, and decreased by DNp63b overexpression, Wnt5a knockdown, or Ror2 knockdown.

Article Snippet: Exogenous Wnt5a treatment Recombinant human Wnt5a protein (rhWnt5a) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Over Expression, Knockdown, MANN-WHITNEY, Zymography Assay

Journal: Oral oncology

Article Title: Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition.

doi: 10.1016/j.oraloncology.2017.03.019

Figure Lengend Snippet: Fig. 6. Effects of rhWnt5a protein on migration, invasion, and activity of MMP-2 in tongue SCC cells. Tongue SCC cells were treated with increasing concentrations of rhWnt5a (0, 0.01, 0.1, and 1 lg/mL). (A) Wound healing assay shows that addition of rhWnt5a for 24 h increases migration abilities of SQUU-B cells transfected with Wnt5a siRNA (Dunnett’s test, p < 0.05). Stimulation with rhWnt5a has no effect on the migration abilities of SQUU-A, SQUU-BO, or Ror2-depleted SQUU-B cells. Scale bars, 200 lm (B) Matrigel TM invasion assay, addition of rhWnt5a for 22 h increases the number of invading SQUU-B cells transfected Wnt5a siRNA (Dunnett’s test, p < 0.05). Stimulation with rhWnt5a has no effect on the invasion abilities of SQUU-A, SQUU-BO, or Ror2-depleted SQUU-B cells. Scale bars, 100 lm.(C) Gelatin zymography assay shows that preformed and active MMP-2 are increased in Wnt5a-depleted SQUU-B cells following stimulation with rhWnt5a.

Article Snippet: Exogenous Wnt5a treatment Recombinant human Wnt5a protein (rhWnt5a) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Migration, Activity Assay, Wound Healing Assay, Transfection, Invasion Assay, Zymography Assay

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: Figure 1. 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

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: Figure 2. 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.

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

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: Figure 3. 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.

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

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: Figure 4. 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.

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: Expressing, Cell Culture, Isolation, Quantitative RT-PCR, Gene Expression

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: Figure 5. 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.

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, Western Blot

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: Figure 7. 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.

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: Cell Culture, Isolation, Quantitative RT-PCR, Expressing, Gene Expression

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: Figure 8. 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.

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: Cell Culture, 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: Figure 9. 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.

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: Expressing, Immunohistochemical staining, Staining

Journal: Advanced Science

Article Title: Single‐Cell RNA Sequencing Identifies MMP11 + Cancer‐Associated Fibroblasts as Drivers of Angiogenesis and Bladder Cancer Progression

doi: 10.1002/advs.202502774

Figure Lengend Snippet: MMP11 + mCAFs drive the migration of ESM1 + tECs through the WNT5A‐MCAM signaling axis. A) Signaling output landscape of different cell populations, visualized to highlight key pathways and intercellular communication hubs. B) Communication intensity of the WNT5A‐MCAM signaling axis between MMP11 + mCAFs with distinct cell populations. C) UMAP plot displaying the classification of endothelial cell subpopulation. D) Expression levels of MCAM across identified endothelial subpopulations, visualized as a UMAP plot (top) and a Dot plot (bottom) overlay. E) GO enrichment analysis showing key biological processes and pathways associated with upregulated genes in ESM1 + tECs. F) Heatmap of pathway activity variations scored by GSVA for each cell between different endothelial groups. G) IF staining for spatial localization of MMP11 + mCAFs, confirming their proximity to endothelial regions of interest. Scale bar = 20 µm. The white arrow indicates the MMP11 + mCAF cells. H) Transwell migration assay demonstrating the effect of WNT5A and Box5‐TFA, with representative images showing cell migration trends. Scale bar = 50 µm, n = 3. I) Angiogenesis assays assessing effect of WNT5A and Box5‐TFA on tube formation activity in HUVECs. Scale bar = 50 µm, n = 3. J–L) Quantifications of migrated cells in Transwell migration assay (J), junction numbers (K), and mesh numbers (L) in Angiogenesis assays. Statistical analysis was performed using Student's t ‐test; ** p < 0.01, *** p < 0.001, **** p < 0.0001. M) Transcription factors specific to endothelial subpopulations, identified via single‐cell transcriptomics. N) ESM1 + tEC‐specific transcription factors and their regulatory target genes, mapped to demonstrate transcriptional control over ESM1 + tEC signature genes. O) Correlation between ESM1 + tEC‐specific transcription factors genes and MCAM expression, tested by Spearman correlation.

Article Snippet: The treatments included 10 ng mL −1 WNT5A (Yeasen, Shanghai, China; 92282ES10), 10 μ m Box5‐TFA (MedChemExpress, New Jersey, USA; HY‐123071A), a combination of the two agents, and PBS as a control.

Techniques: Migration, Expressing, Activity Assay, Staining, Transwell Migration Assay, Single-cell Transcriptomics, Control