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wnt5a  (Cusabio)


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    Cusabio wnt5a
    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, <t>WNT5A</t> , 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.
    Wnt5a, supplied by Cusabio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+wnt5a/Recombinant+Human+Protein+Wnt-5a%EF%BC%88WNT5A%EF%BC%89/pmc12444465-490-13-17
    Average 93 stars, based on 1 article reviews
    wnt5a - by Bioz Stars, 2026-10
    93/100 stars

    Images

    1) Product Images from "Microfluidic chip-integrated vascularized endometrial complexes: Mitochondrial function and paracrine crosstalk enhance regenerative potential"

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

    Journal: Bioactive Materials

    doi: 10.1016/j.bioactmat.2025.08.035

    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.
    Figure Legend 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.

    Techniques Used: 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.
    Figure Legend 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 Used: Protein-Protein interactions, Cell Culture, Control, Positive Control, Software

    Related Articles

    Immunohistochemistry:

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517–787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811–2910).

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer.
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/ DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. 1B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517– 787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811– 2910).

    Bicinchoninic Acid Protein Assay:

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517–787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811–2910).

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer.
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/ DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. 1B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517– 787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811– 2910).

    Recombinant:

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517–787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811–2910).

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer.
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/ DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. 1B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517– 787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811– 2910).

    Cell Counting:

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517–787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811–2910).

    Article Title: ATBF1 is a potential diagnostic marker of histological grade and functions via WNT5A in breast cancer.
    Article Snippet: .. The reagents were purchased from their respective vendors: expose mouse- and rabbit-specific HRP/ DAB detection IHC kit (Cat. ab236466, Abcam, Shanghai, China); RNA-Solv Reagent (Cat. R6830, Omega, Guangzhou, China); TRIzolTM (Cat. 15,596,026, Invitrogen, Shanghai, China); BCA Protein Assay Kit (Cat. CW0014S, CWBIO, Jiangsu, China); secondary antibodies (BOSTER, Wuhan, China); Lipofectamine RNAiMAX reagent (Cat. 13,778,100, Invitrogen); recombinant human WNT5A (Cat. CSB-EP026138HU1, CUSABIO, Wuhan, China); cell counting kit (APE×Bio, Shanghai, China); WNT5A primary antibody (Cat. A19133, Abclonal, Wuhan, China); and β-actin primary antibody (Cat. AC026, Abclonal). .. ATBF1 primary antibodies were designed to recognize independent peptides, as shown in Fig. 1B: BC029653 (Abcam, Shanghai, China), which recognizes the N-terminus of ATBF1 (amino acids, 517– 787); D1-120 (MBL, Shanghai, China), which recognizes a middle region of ATBF1 (amino acids, 2107–2147); and 3B1 (Santa Cruz Biotechnology, Shanghai, China), which recognizes the C-terminus of ATBF1 (amino acids, 2811– 2910).



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    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, <t>WNT5A</t> , 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.
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    ( A ) UMAP of epithelial subtypes in control and NEC mice. ( B ) Representative immunofluorescence images for Lyz (red), Epacm (green) and DAPI (blue) on paraffin-embedded distal ileum from indicated group in experiment 3. White arrow indicates the Lyz stained PCs, Scale bar: 10 μm. ( C , D ) The number of Lyz + PCs in B ( C ) and mRNA levels of Lyz ( D ) were compared among different groups. ( E ) Violin plots showed the expression of Wnt/PCP pathway genes in mouse PCs subsets revealed by scRNA-seq analysis. ( F ) The mRNA level of Damm1 was compared in IECs isolated from different groups. ( G ) Organoids derived from mice were cultured in medium with DMSO, FEX (5 μm), or FEX (10 μm) from day 1. The organoids were collected and stained for Lyz on day 7. Arrowheads point at PCs (Lyz + , in pink), scale bars: 50 μm. ( H ) qRT-PCR analysis of Daam1 in organoids cultured with DMSO, FEX (5 μm), or FEX (10 μm). ( I ) Representative organoid formation images on day 7 in different groups supplemented with ENR, Wnt5a-containing ENR, and Wnt5a-containing ENR + FEX. Representative marker genes for PCs (Lyz, green) cells are highlighted by fluorescent reporters. Arrowheads point at PCs (Lyz + , in green), scale bars: 50 μm. All statistical data are shown as the means ± SEMs, *p < 0.05, **p < 0.01, ***p < 0.001.

    Journal: NPJ Biofilms and Microbiomes

    Article Title: Disruption of bile acid homeostasis potentiates Paneth cell ablation by activating the intestinal Farnesoid X receptor in necrotizing enterocolitis

    doi: 10.1038/s41522-025-00904-6

    Figure Lengend Snippet: ( A ) UMAP of epithelial subtypes in control and NEC mice. ( B ) Representative immunofluorescence images for Lyz (red), Epacm (green) and DAPI (blue) on paraffin-embedded distal ileum from indicated group in experiment 3. White arrow indicates the Lyz stained PCs, Scale bar: 10 μm. ( C , D ) The number of Lyz + PCs in B ( C ) and mRNA levels of Lyz ( D ) were compared among different groups. ( E ) Violin plots showed the expression of Wnt/PCP pathway genes in mouse PCs subsets revealed by scRNA-seq analysis. ( F ) The mRNA level of Damm1 was compared in IECs isolated from different groups. ( G ) Organoids derived from mice were cultured in medium with DMSO, FEX (5 μm), or FEX (10 μm) from day 1. The organoids were collected and stained for Lyz on day 7. Arrowheads point at PCs (Lyz + , in pink), scale bars: 50 μm. ( H ) qRT-PCR analysis of Daam1 in organoids cultured with DMSO, FEX (5 μm), or FEX (10 μm). ( I ) Representative organoid formation images on day 7 in different groups supplemented with ENR, Wnt5a-containing ENR, and Wnt5a-containing ENR + FEX. Representative marker genes for PCs (Lyz, green) cells are highlighted by fluorescent reporters. Arrowheads point at PCs (Lyz + , in green), scale bars: 50 μm. All statistical data are shown as the means ± SEMs, *p < 0.05, **p < 0.01, ***p < 0.001.

    Article Snippet: For experiments involving Wnt5a (645-WN-010/CF, R&D Systems) ligands, the media was made by supplementing ENR media with 100 ng/ml Wnt5a.

    Techniques: Control, Immunofluorescence, Staining, Expressing, Isolation, Derivative Assay, Cell Culture, Quantitative RT-PCR, Marker

    a General schematic of the TetON inducible gene expression system used in this study. In the absence of doxycycline, the reverse tetracycline-controlled transactivator (rtTA) is inactive. Upon doxycycline addition, rtTA binds to the tetO promoter and activates transcription of the gene of interest, leading to protein expression. b Doxycycline-induced overexpression of HA-tagged PRICKLE3 in HEK T-REx 293 PRICKLE3 TetON cells and of PRICKLE1 in the corresponding inducible cell line. c Induction of HA-PRICKLE3 expression by doxycycline in HEK T-REx 293 PRICKLE3 TetON cells. HEK T-REx 293 wildtype (WT) cells served as a control for doxycycline effects. Arrowheads indicate phosphorylation-dependent shifts in the electrophoretic mobility of VANGL proteins. α-TUBULIN was used as a loading control. Representative result from n = 5. d , e Densitometric quantification of Western blot signals. Values were normalized to untreated cells. Statistical analysis was performed using an unpaired t -test; corresponding p -values are shown ( n = 5). f Effect of recombinant WNT stimulation on VANGL phosphorylation. HEK T-REx 293 PRICKLE3 TetON cells were pre-treated overnight with the porcupine inhibitor LGK-974 to block endogenous WNT ligand secretion and subsequently stimulated with 100 ng/ml human recombinant WNT5A or WNT3A for 3 h. Arrowheads indicate phosphorylation-dependent mobility shifts of ROR. α-TUBULIN served as a loading control. Representative result from n = 4. g , h Densitometric quantification of Western blot signals from panel f. Data were normalized to untreated controls. Statistical analysis was performed using an unpaired t -test; corresponding p -values are reported ( n = 4).

    Journal: Communications Biology

    Article Title: PRICKLE3 protects VANGL proteins from CK1-mediated phosphorylation and RNF43-mediated degradation

    doi: 10.1038/s42003-025-09422-9

    Figure Lengend Snippet: a General schematic of the TetON inducible gene expression system used in this study. In the absence of doxycycline, the reverse tetracycline-controlled transactivator (rtTA) is inactive. Upon doxycycline addition, rtTA binds to the tetO promoter and activates transcription of the gene of interest, leading to protein expression. b Doxycycline-induced overexpression of HA-tagged PRICKLE3 in HEK T-REx 293 PRICKLE3 TetON cells and of PRICKLE1 in the corresponding inducible cell line. c Induction of HA-PRICKLE3 expression by doxycycline in HEK T-REx 293 PRICKLE3 TetON cells. HEK T-REx 293 wildtype (WT) cells served as a control for doxycycline effects. Arrowheads indicate phosphorylation-dependent shifts in the electrophoretic mobility of VANGL proteins. α-TUBULIN was used as a loading control. Representative result from n = 5. d , e Densitometric quantification of Western blot signals. Values were normalized to untreated cells. Statistical analysis was performed using an unpaired t -test; corresponding p -values are shown ( n = 5). f Effect of recombinant WNT stimulation on VANGL phosphorylation. HEK T-REx 293 PRICKLE3 TetON cells were pre-treated overnight with the porcupine inhibitor LGK-974 to block endogenous WNT ligand secretion and subsequently stimulated with 100 ng/ml human recombinant WNT5A or WNT3A for 3 h. Arrowheads indicate phosphorylation-dependent mobility shifts of ROR. α-TUBULIN served as a loading control. Representative result from n = 4. g , h Densitometric quantification of Western blot signals from panel f. Data were normalized to untreated controls. Statistical analysis was performed using an unpaired t -test; corresponding p -values are reported ( n = 4).

    Article Snippet: Following this, non-canonical WNT signaling was activated by incubating the cells with recombinant human WNT5A (645-WN, R&D Systems) at a concentration of 100 ng/mL for 3 h. Similarly, canonical WNT signaling was activated by incubating the cells with 100 ng/mL of recombinant human WNT3A (5036-WN, R&D Systems) for 3 h. To assess protein degradation kinetics, cells were pretreated with doxycycline overnight.

    Techniques: Gene Expression, Expressing, Over Expression, Control, Phospho-proteomics, Western Blot, Recombinant, Blocking Assay

    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.

    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