wnt16  (Proteintech)

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: WNT16 is highly expressed in DSCs of normal early pregnancy. (A) Venn diagram depicting 526 genes upregulated during physiological decidualization in normal pregnancy from two datasets. (B) KEGG enrichment analysis was performed on the intersecting genes. (C) GSEA of the WNT signaling pathway based on the whole-gene ranked lists from the comparisons of dESCs vs ESCs, and RSA-DSCs vs NP-DSCs. (D) Heatmap showing the RT-qPCR expression patterns of WNT ligands across ESCs, NP-DSCs, and RSA-DSCs. (E) Bar plot displaying the log 2 fold changes (log 2 FC) in WNT ligand expression (left: NP-DSCs vs ESCs; right: RSA-DSCs vs NP-DSCs). (F) Representative immunohistochemical showing WNT16 expression in endometrial from non-pregnancy women and in decidual tissues from NP and RSA pregnancies (n=4), together with quantitative analysis (G) . (H) ELISA quantification of WNT16 in supernatants from 72h cultures of ESCs, NP-DSCs, and RSA-DSCs (n=4). Data are presented as mean ± SEM. **p < 0.01, ***p < 0.001.

Article Snippet: The membrane was blocked with 5% skim milk for 1h at room temperature and incubated with primary antibodies against β-catenin (1:15000, ProteinTech, USA) or WNT16 (1:4000, ProteinTech, USA) overnight at 4 °C.

Techniques: Quantitative RT-PCR, Expressing, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: WNT16 drives toward an M2-like phenotype. (A) Volcano plot of DEGs in WNT16-treated pMo relative to untreated controls(∣log 2 FC∣> 1 and P< 0.05). (B) GO analysis of DEGs showing the top 10 enriched biological process terms, ranked by adjusted P value. (C) GSEA of immune-related pathways based on DEGs. (D) FCM analysis of representative markers in pMo after 48 hours of WNT16 treatment (n=3-5). (E) ELISA measurement of IL-10 and IFN-γ levels in culture supernatants from WNT16-treated pMo (n=3). Data are presented as mean ± SEM. NS, not significant; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The membrane was blocked with 5% skim milk for 1h at room temperature and incubated with primary antibodies against β-catenin (1:15000, ProteinTech, USA) or WNT16 (1:4000, ProteinTech, USA) overnight at 4 °C.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: DSC-derived WNT16 activates canonical WNT signaling pathway in dMφ. (A) GSEA using DEGs from WNT16-treated versus control pMo, ranked by absolute NES score (|NES|). (B-C) Heatmap comparing mRNA expression levels of WNT receptor family members between pMo and dMφ (B) , with corresponding quantitative expression levels represented as log 2 FC in bar plots (C) . (D-E) Western Blot analysis of β-catenin in dMφ after WNT16 treatment (D) or with various concentrations of DSC-CM (E) , with corresponding quantification shown as bar graphs (right panels) (n=3). (F) Representative immunofluorescence images showing β-catenin nuclear localization in dMφ; scale bar: 20 μm. (G) TOP-flash and FOP-flash luciferase activity measured in dMφ after WNT16 treatment (n=3). Data are presented as mean ± SEM. NS, not significant; **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 1h at room temperature and incubated with primary antibodies against β-catenin (1:15000, ProteinTech, USA) or WNT16 (1:4000, ProteinTech, USA) overnight at 4 °C.

Techniques: Derivative Assay, Control, Expressing, Western Blot, Immunofluorescence, Luciferase, Activity Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: WNT16 remodels chromatin accessibility to promote an M2-polarized epigenetic landscape. (A) Heatmaps of chromatin accessibility in regions ±3 kb around TSS. (B) Genomic distribution of transcription factor binding sites in relation to TSS. (C) Distribution of accessible chromatin across various genomic features. GO (D) and KEGG (E) enrichment analysis of genes with upregulated chromatin accessibility in dMφ after WNT16 stimulation. (F) Gene tracks from ATAC-seq showing chromatin accessibility at representative WNT signaling genes ( CTNNB1 , AXIN2 ) and M2 macrophage polarization markers ( CD163 , IL-10 ) in dMφ with or without WNT16 treatment. (G) Top six enriched TF motifs identified in WNT16-treated dMφ by HOMER motif analysis.

Article Snippet: The membrane was blocked with 5% skim milk for 1h at room temperature and incubated with primary antibodies against β-catenin (1:15000, ProteinTech, USA) or WNT16 (1:4000, ProteinTech, USA) overnight at 4 °C.

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: Integrated multi-omics analysis of a MYC–DIXDC1 axis in relation to Wnt/β-catenin signaling in M2 dMφ. (A) Venn diagram showing 13 genes commonly upregulated in both ATAC-seq and RNA-seq analyses after WNT16 treatment. (B) Volcano plot displays these overlapping DEGs. (C) IGV tracks of three M2 polarization-associated genes selected from the 13 overlapping upregulated genes. (D) KEGG pathway analysis of the 13 overlapping upregulated DEGs. (E) IGV tracks illustrating chromatin accessibility of DIXDC1. (F) Venn diagram showing transcription factors predicted to regulate DIXDC1 across eight databases. (G) Intersection of predicted transcription factors with TFs upregulated in ATAC-seq and RNA-seq analyses. (H) The relative expression levels of macrophage polarization markers in dMφ were determined by RT-qPCR after the treatment with siDIXDC1, WNT16 or siDIXDC1 plus WNT16 (n=3). (I) Western blot analysis of β-catenin and WNT16 protein expressions under the same experimental conditions. Data are presented as mean ± SEM. NS, not significant; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The membrane was blocked with 5% skim milk for 1h at room temperature and incubated with primary antibodies against β-catenin (1:15000, ProteinTech, USA) or WNT16 (1:4000, ProteinTech, USA) overnight at 4 °C.

Techniques: Biomarker Discovery, RNA Sequencing, Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: Schematic diagram of DSC-derived WNT16 promoting M2 dMφ polarization at the maternal-fetal interface. During normal pregnancy, DSCs secrete WNT16 that binds to FZD receptors on dMφ. This triggers the canonical WNT/β-catenin signaling pathway, leading to β-catenin activation and its nuclear translocation. Once in the nucleus, β-catenin may participate in transcriptional regulation together with TCF/LEF factors, potentially initiating the transcription of MYC and several M2 macrophage-associated genes. Subsequently, MYC could contribute to the transcription regulation of DIXDC1 , a known activator of WNT/β-catenin pathway, which may help maintain the M2-associated transcriptional program and support an immunotolerant environment during pregnancy. Created with BioRender.com .

Article Snippet: The membrane was blocked with 5% skim milk for 1h at room temperature and incubated with primary antibodies against β-catenin (1:15000, ProteinTech, USA) or WNT16 (1:4000, ProteinTech, USA) overnight at 4 °C.

Techniques: Derivative Assay, Activation Assay, Translocation Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: WNT16 is highly expressed in DSCs of normal early pregnancy. (A) Venn diagram depicting 526 genes upregulated during physiological decidualization in normal pregnancy from two datasets. (B) KEGG enrichment analysis was performed on the intersecting genes. (C) GSEA of the WNT signaling pathway based on the whole-gene ranked lists from the comparisons of dESCs vs ESCs, and RSA-DSCs vs NP-DSCs. (D) Heatmap showing the RT-qPCR expression patterns of WNT ligands across ESCs, NP-DSCs, and RSA-DSCs. (E) Bar plot displaying the log 2 fold changes (log 2 FC) in WNT ligand expression (left: NP-DSCs vs ESCs; right: RSA-DSCs vs NP-DSCs). (F) Representative immunohistochemical showing WNT16 expression in endometrial from non-pregnancy women and in decidual tissues from NP and RSA pregnancies (n=4), together with quantitative analysis (G) . (H) ELISA quantification of WNT16 in supernatants from 72h cultures of ESCs, NP-DSCs, and RSA-DSCs (n=4). Data are presented as mean ± SEM. **p < 0.01, ***p < 0.001.

Article Snippet: Sections were incubated overnight with WNT16 antibody (1:200, ProteinTech, USA) in a humidity chamber.

Techniques: Quantitative RT-PCR, Expressing, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: WNT16 drives toward an M2-like phenotype. (A) Volcano plot of DEGs in WNT16-treated pMo relative to untreated controls(∣log 2 FC∣> 1 and P< 0.05). (B) GO analysis of DEGs showing the top 10 enriched biological process terms, ranked by adjusted P value. (C) GSEA of immune-related pathways based on DEGs. (D) FCM analysis of representative markers in pMo after 48 hours of WNT16 treatment (n=3-5). (E) ELISA measurement of IL-10 and IFN-γ levels in culture supernatants from WNT16-treated pMo (n=3). Data are presented as mean ± SEM. NS, not significant; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Sections were incubated overnight with WNT16 antibody (1:200, ProteinTech, USA) in a humidity chamber.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: DSC-derived WNT16 activates canonical WNT signaling pathway in dMφ. (A) GSEA using DEGs from WNT16-treated versus control pMo, ranked by absolute NES score (|NES|). (B-C) Heatmap comparing mRNA expression levels of WNT receptor family members between pMo and dMφ (B) , with corresponding quantitative expression levels represented as log 2 FC in bar plots (C) . (D-E) Western Blot analysis of β-catenin in dMφ after WNT16 treatment (D) or with various concentrations of DSC-CM (E) , with corresponding quantification shown as bar graphs (right panels) (n=3). (F) Representative immunofluorescence images showing β-catenin nuclear localization in dMφ; scale bar: 20 μm. (G) TOP-flash and FOP-flash luciferase activity measured in dMφ after WNT16 treatment (n=3). Data are presented as mean ± SEM. NS, not significant; **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Sections were incubated overnight with WNT16 antibody (1:200, ProteinTech, USA) in a humidity chamber.

Techniques: Derivative Assay, Control, Expressing, Western Blot, Immunofluorescence, Luciferase, Activity Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: WNT16 remodels chromatin accessibility to promote an M2-polarized epigenetic landscape. (A) Heatmaps of chromatin accessibility in regions ±3 kb around TSS. (B) Genomic distribution of transcription factor binding sites in relation to TSS. (C) Distribution of accessible chromatin across various genomic features. GO (D) and KEGG (E) enrichment analysis of genes with upregulated chromatin accessibility in dMφ after WNT16 stimulation. (F) Gene tracks from ATAC-seq showing chromatin accessibility at representative WNT signaling genes ( CTNNB1 , AXIN2 ) and M2 macrophage polarization markers ( CD163 , IL-10 ) in dMφ with or without WNT16 treatment. (G) Top six enriched TF motifs identified in WNT16-treated dMφ by HOMER motif analysis.

Article Snippet: Sections were incubated overnight with WNT16 antibody (1:200, ProteinTech, USA) in a humidity chamber.

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: Integrated multi-omics analysis of a MYC–DIXDC1 axis in relation to Wnt/β-catenin signaling in M2 dMφ. (A) Venn diagram showing 13 genes commonly upregulated in both ATAC-seq and RNA-seq analyses after WNT16 treatment. (B) Volcano plot displays these overlapping DEGs. (C) IGV tracks of three M2 polarization-associated genes selected from the 13 overlapping upregulated genes. (D) KEGG pathway analysis of the 13 overlapping upregulated DEGs. (E) IGV tracks illustrating chromatin accessibility of DIXDC1. (F) Venn diagram showing transcription factors predicted to regulate DIXDC1 across eight databases. (G) Intersection of predicted transcription factors with TFs upregulated in ATAC-seq and RNA-seq analyses. (H) The relative expression levels of macrophage polarization markers in dMφ were determined by RT-qPCR after the treatment with siDIXDC1, WNT16 or siDIXDC1 plus WNT16 (n=3). (I) Western blot analysis of β-catenin and WNT16 protein expressions under the same experimental conditions. Data are presented as mean ± SEM. NS, not significant; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Sections were incubated overnight with WNT16 antibody (1:200, ProteinTech, USA) in a humidity chamber.

Techniques: Biomarker Discovery, RNA Sequencing, Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Immunology

Article Title: WNT16 from decidual stromal cells orchestrates M2 macrophage polarization via β-catenin signaling and chromatin remodeling at the maternal-fetal interface

doi: 10.3389/fimmu.2025.1712898

Figure Lengend Snippet: Schematic diagram of DSC-derived WNT16 promoting M2 dMφ polarization at the maternal-fetal interface. During normal pregnancy, DSCs secrete WNT16 that binds to FZD receptors on dMφ. This triggers the canonical WNT/β-catenin signaling pathway, leading to β-catenin activation and its nuclear translocation. Once in the nucleus, β-catenin may participate in transcriptional regulation together with TCF/LEF factors, potentially initiating the transcription of MYC and several M2 macrophage-associated genes. Subsequently, MYC could contribute to the transcription regulation of DIXDC1 , a known activator of WNT/β-catenin pathway, which may help maintain the M2-associated transcriptional program and support an immunotolerant environment during pregnancy. Created with BioRender.com .

Article Snippet: Sections were incubated overnight with WNT16 antibody (1:200, ProteinTech, USA) in a humidity chamber.

Techniques: Derivative Assay, Activation Assay, Translocation Assay