ha tag  (Sino Biological)

Journal: Communications Biology

Article Title: Wnt11 mediates fibroblast–smooth muscle cell interaction to promote neurogenic bladder fibrosis in rats

doi: 10.1038/s42003-026-09647-2

Figure Lengend Snippet: a Relative mRNA level of WNT11 in fibrotic bladders following BPNI and SCI ( n = 8). Western blotting of WNT11 expression in BPNI ( b ) and SCI ( c )-induced NB. Representative photomicrographs showed that WNT11 (green) was increased in rats after BPNI ( d ) and SCI ( e ). Scale bar = 400 μm. Protein levels of fibroblasts differentiation-related markers (α-SMA and Col-1) in BFs isolated from rats treated with BPNI ( f ) and SCI ( g ). Representative western blotting showing the phenotypic transformation of SMCs (SMTN and MYH10) in BPNI ( h ) and SCI ( i ) rats. Western blotting analyses showing protein levels of WNT11 and markers of fibroblastic differentiation (α-SMA and Col-1) and SMCs phenotypic transformation (SMTN and MYH10) in BFs ( j ) and SMCs ( k ) treated with TGF-β1 at different doses, respectively. Data are shown as the mean ± SD, and P values were determined by the one-way ANOVA followed by Tukey’s post-hoc test ( a ), * P < 0.05, ** P < 0.01.

Article Snippet: To maximally activate or inhibit the WNT cascade in vivo, rats were treated with recombinant WNT11 protein (#6179-WN-010, R&D Systems) or LGK974 (#HY-17545, MedChemExpress) immediately after the surgical procedure.

Techniques: Western Blot, Expressing, Isolation, Transformation Assay

Journal: Communications Biology

Article Title: Wnt11 mediates fibroblast–smooth muscle cell interaction to promote neurogenic bladder fibrosis in rats

doi: 10.1038/s42003-026-09647-2

Figure Lengend Snippet: a – n Eight-week-old male Sprague–Dawley rats were subjected to sham, BPNI, or SCI operation. Vehicle, WNT11, or LGK974 were administered to rats every day after the surgical procedure for 2 weeks, and these rats were euthanized at post 4 weeks of operation. a Schematic of the experimental protocol and treatment regimen. b Representative bladder images in rats treated with vehicle, WNT11, or LGK974 post BPNI or SCI. Representative images of cystometrograms in rats after BPNI ( c ) or SCI ( d ). Statistical results for the intravesical pressure in rats after BPNI ( e ) or SCI ( f ) ( n = 5). Histological analysis of the Masson’s trichrome staining in bladders after BPNI ( g ) or SCI ( h ). Scale bar = 200 μm. Quantifications of detrusor thickness ( i , j ) and collagen content ( k , l ) by Masson’s trichrome staining were performed to assess the bladder fibrosis ( n = 5). Western blotting showing the protein levels of WNT11 and markers of fibroblastic differentiation (α-SMA and Col-1) and SMCs phenotypic transformation (SMTN and MYH10) in bladders after BPNI ( m ) or SCI ( n ). Western blotting analyses showing protein levels of TGF-β1 and markers of fibroblastic differentiation (α-SMA and Col-1) and SMCs phenotypic transformation (SMTN and MYH10) in BFs ( o ) and SMCs ( p ) treated with WNT11 at different doses. Western blotting analyses showing siWNT11 transfection inhibiting fibroblastic differentiation and SMCs phenotypic transformation in BFs ( q ) and SMCs ( r ) with TGF-β1 treatment, respectively. Data are shown as the mean ± SD, and P values were determined by the two-way ANOVA followed by Tukey’s post-hoc test ( e , f , i – l ), ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: To maximally activate or inhibit the WNT cascade in vivo, rats were treated with recombinant WNT11 protein (#6179-WN-010, R&D Systems) or LGK974 (#HY-17545, MedChemExpress) immediately after the surgical procedure.

Techniques: Staining, Western Blot, Transformation Assay, Transfection

Journal: Communications Biology

Article Title: Wnt11 mediates fibroblast–smooth muscle cell interaction to promote neurogenic bladder fibrosis in rats

doi: 10.1038/s42003-026-09647-2

Figure Lengend Snippet: Representative western blotting and quantitative analyses for FZD10 and Vangl2 in bladders after BPNI ( a ) or SCI ( b ) ( n = 6). Interaction between endogenous WNT11 and Vangl2 in BFs ( c ) and SMCs ( d ). e Confocal microscopy analysis of BFs and SMCs showing the colocalization of WNT11 (red) and Vangl2 (green). Scale bar = 20 μm. Quantification of colocalization was ascertained by ImageJ software in five randomly chosen fields. Numbers in the images correspond to the average Pearson’s correlation coefficient ± SD. Representative Western blotting and summarized data showing the relative protein levels of Vangl2 and markers of fibroblastic differentiation (α-SMA and Col-1) and SMCs phenotypic transformation (SMTN and MYH10) in BFs ( f ) and SMCs ( g ) ( n = 4). Data are shown as the mean ± SD. P values were determined by the two-tailed unpaired Student’s t -test ( a , b ) and the two-way ANOVA followed by Tukey’s post-hoc test ( f , g ), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: To maximally activate or inhibit the WNT cascade in vivo, rats were treated with recombinant WNT11 protein (#6179-WN-010, R&D Systems) or LGK974 (#HY-17545, MedChemExpress) immediately after the surgical procedure.

Techniques: Western Blot, Confocal Microscopy, Software, Transformation Assay, Two Tailed Test

Journal: Communications Biology

Article Title: Wnt11 mediates fibroblast–smooth muscle cell interaction to promote neurogenic bladder fibrosis in rats

doi: 10.1038/s42003-026-09647-2

Figure Lengend Snippet: Representative western blotting and quantitative analyses for the phosphorylation levels of JNK and c-JUN in bladders after BPNI ( a ) or SCI ( b ) ( n = 3). BFs ( c ) and SMCs ( d ) developed higher expression levels of p-JNK and p-c-JUN upon WNT11 treatment as indicated by the western blotting and quantitative analyses ( n = 4). Representative immunofluorescence staining and quantitative analyses showing that WNT11 treatment increased the fluorescence intensity and accumulation of p-c-JUN in the nucleus of BFs ( e ) and SMCs ( f ) ( n = 4). Scale bar=100μm. Protein levels of p-JNK, p-c-JUN, and markers of fibroblastic differentiation (α-SMA and Col-1) and SMCs phenotypic transformation (SMTN and MYH10) in BFs ( g ) and SMCs ( h ) treated with WNT11 with or without SP600125 by western blotting ( n = 3). Data are shown as the mean ± SD. P values were determined by the two-way ANOVA followed by Tukey’s post-hoc test ( a , b , g , h ) and the two-tailed unpaired student’s t -test ( c – f ), * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To maximally activate or inhibit the WNT cascade in vivo, rats were treated with recombinant WNT11 protein (#6179-WN-010, R&D Systems) or LGK974 (#HY-17545, MedChemExpress) immediately after the surgical procedure.

Techniques: Western Blot, Phospho-proteomics, Expressing, Immunofluorescence, Staining, Fluorescence, Transformation Assay, Two Tailed Test

Journal: Communications Biology

Article Title: Wnt11 mediates fibroblast–smooth muscle cell interaction to promote neurogenic bladder fibrosis in rats

doi: 10.1038/s42003-026-09647-2

Figure Lengend Snippet: Relative mRNA levels of DVL1, DVL2, and DVL3 in fibrotic bladders at 4 weeks after BPNI ( a ) and SCI ( b ) ( n = 6). Representative western blotting and quantitative analyses for the protein levels of DVL1, DVL2, and DVL3 in bladders after BPNI ( c ) or SCI ( d ) ( n = 6). e Representative western blotting and quantitative analyses revealing the protein levels of DVL2 and markers of fibroblastic differentiation, and the phosphorylation levels of JNK and c-JUN in BFs ( n = 3). f Protein levels of p-c-JUN, p-JNK, DVL2, SMTN, and MYH10 in SMCs treated with WNT11 and/or siDVL2 ( n = 3). Immunoprecipitation demonstrated that DVL2 bound to Vangl2 in BFs ( g ) and SMCs ( h ). Representative western blotting showing the presence of the target proteins (input) and immunoprecipitation showing the interaction of Flag-DVL2, Flag-DEP, Flag-DIX, and Flag-PDZ with endogenous Vangl2 in HEK293T cells using antibodies to Flag ( i ) and IgG ( j ). Data are shown as the mean ± SD. P values were determined by the two-tailed unpaired Student’s t -test ( a – d ) and the two-way ANOVA followed by Tukey’s post-hoc test ( e , f ), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: To maximally activate or inhibit the WNT cascade in vivo, rats were treated with recombinant WNT11 protein (#6179-WN-010, R&D Systems) or LGK974 (#HY-17545, MedChemExpress) immediately after the surgical procedure.

Techniques: Western Blot, Phospho-proteomics, Immunoprecipitation, Two Tailed Test

Journal: Communications Biology

Article Title: Wnt11 mediates fibroblast–smooth muscle cell interaction to promote neurogenic bladder fibrosis in rats

doi: 10.1038/s42003-026-09647-2

Figure Lengend Snippet: Under the condition of bladder denervation, the WNT11/Vangl2 signal was transmitted into the cytoplasm via DVL2, followed by activating PCP signaling to promote fibroblastic differentiation and SMC phenotypic transformation, which also depended on the interaction of TGF-β1/Smad2/3 signaling at the level of cytomembrane and nucleus, eventually resulting in NB fibrosis.

Article Snippet: To maximally activate or inhibit the WNT cascade in vivo, rats were treated with recombinant WNT11 protein (#6179-WN-010, R&D Systems) or LGK974 (#HY-17545, MedChemExpress) immediately after the surgical procedure.

Techniques: Transformation Assay

Journal: Scientific Reports

Article Title: Differential gene expression in trabecular bone osteocytes is related to the local strain and strain gradient

doi: 10.1038/s41598-025-00214-z

Figure Lengend Snippet: ROCs indicate that deviatoric strain and hydrostatic strain gradient were both relatively weak discriminators of altered TNF ( a ) and FRZB ( b ) expression. Altered SOST expression was better identified by strain gradient ( c ), while Wnt11 expression was similarly discriminated by each measure ( d ). Excluding the osteocytes where only one gene was altered increased the AUC in all cases. ( e ) Hydrostatic strain gradient was a better predictor than deviatoric strain for combined SOST and Wnt11 expression than deviatoric strain. ( f ) Deviatoric strain was a better predictor of TNF + FRZB expression than hydrostatic strain gradient.

Article Snippet: Custom probes for porcine TNF, FRZB, SOST, and Wnt11 were designed and obtained from ACDBio (Supplemental Table ).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Non-Canonical Wnt11 Signaling Regulates Pulmonary Fibrosis via Fibroblast and Alveolar Epithelial Type II Cell Crosstalk

doi: 10.3390/ijms27010351

Figure Lengend Snippet: Wnt11 induction in human lung fibroblast is β-catenin dependent. ( A ) Lung tissue RNA from PBS- or BLM-treated mice at day 7 were analyzed for Wnt component mRNA N = 5. ( B ) Human lung fibroblasts (HLFs) were pretreated with GSKβ inhibitor CHIR (5 µM) or solvent DMSO (None) for 24 h, followed by treatment with TGFβ (5 ng/mL) or PBS (None) for 24 h. The cells were harvested after the indicated hours and analyzed for their mRNA level of AXIN2, WNT5a and WNT11 N = 3–6. HLFs were transfected with control or β-catenin (CTNNB1) siRNA (50 nM) for 24 h and then treated with TGFβ (5 ng/mL) or CHIR99021 (CHIR, 5 µM) for 48 h. Total RNA from HLF was analyzed by qPCR to determine the mRNA levels of AXIN2 ( C ), WNT5A and WNT11 ( D ). ( E ) HLFs were treated with TGFβ (5 ng/mL) or PBS (None) for 24 h and total RNA was analyzed by qPCR to determine the mRNA levels of WNT11, ACTA2 and AXIN2. Numbers above the bars indicate fold-change from untreated cells. N = 3–6. p < * 0.05, ** 0.01, *** 0.001, **** 0.0001 between the indicated two groups by one-way ANOVA plus Tukey’s or Student’s t -test.

Article Snippet: Six-week-old female C57BL/6, Wnt11 fl/fl (B6;129-Wnt11tm.1Ser/J, Stock# 030051) and Col-Cre +/− (C57BL/6J-Tg[Col1α2-Cre-ER(T)], stock# 029567) mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Solvent, Transfection, Control

Journal: International Journal of Molecular Sciences

Article Title: Non-Canonical Wnt11 Signaling Regulates Pulmonary Fibrosis via Fibroblast and Alveolar Epithelial Type II Cell Crosstalk

doi: 10.3390/ijms27010351

Figure Lengend Snippet: Mesenchymal-specific Wnt11 deficiency impaired pulmonary fibrosis and myofibroblast differentiation. Lung fibrosis was induced by bleomycin (BLM) administration following 7 days of tamoxifen injections. ( A ) Representative Trichrome-stained lung tissue sections are shown (scale bar: 500 μm), and Trichome staining was quantified using the Ashcroft scoring method. ( B ) Lung collagen I expression was analyzed at mRNA and protein levels by qPCR ( B ) and Western blotting with quantification ( C ). ( D ) Lung tissue from WT or Wnt11 fl/fl -ColCre +/− cKO mice (cKO) were homogenized at day 21 after BLM treatment and analyzed for collagen content measured by hydroxyproline assay. Mouse lung fibroblasts (MLFs) isolated from BLM-treated WT or cKO mice were further treated with Wnt3a (200 ng/mL) in vitro. MLF RNA collected at 24 h was analyzed for mRNA expression of Wnt11 ( E ) and a-SMA (Acta2) ( F ). MLF lysate collected at 48 h was analyzed for a-SMA protein expression by Western blotting. A representative blot is shown ( G ). N = 3–5. p < * 0.05, ** 0.01, *** 0.001, **** 0.0001 by one-way ANOVA plus Tukey’s or Student’s t -test.

Article Snippet: Six-week-old female C57BL/6, Wnt11 fl/fl (B6;129-Wnt11tm.1Ser/J, Stock# 030051) and Col-Cre +/− (C57BL/6J-Tg[Col1α2-Cre-ER(T)], stock# 029567) mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Staining, Expressing, Western Blot, Hydroxyproline Assay, Isolation, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Non-Canonical Wnt11 Signaling Regulates Pulmonary Fibrosis via Fibroblast and Alveolar Epithelial Type II Cell Crosstalk

doi: 10.3390/ijms27010351

Figure Lengend Snippet: AECII-derived Wnt11 promotes lung myofibroblast differentiation via paracrine signals. ( A ) Primarily isolated mouse AECIIs, A549 cells or ihAEC2s were transfected with the indicated siRNA for 24 h and then treated with TGFβ for another 48 h. After replacing the media, they were co-cultured in Transwell plates with mouse lung fibroblasts (MLFs) or HLF for 48 h. MLFs were harvested and analyzed for mRNA expression of α-SMA (Acta2) and collagen I (Col1a1) by qPCR ( A ). ( B ) AECII RNA samples from ( A ) were analyzed for Wnt11 mRNA by qPCR. The y -axis presents the fold-changes of TGFβ-induced Wnt11 from their untreated samples. ( C ) AECII RNA samples from ( A ) were analyzed for the indicated mRNA by qPCR (fold-change from respective untreated control). ( D ) Primary AECII cultured in 2D on a stiff substrate coated with laminin were collected on day 0 and 3 and analyzed for the indicated mRNA Wnt components by qPCR. ( E ) A549 cells were transfected with WNT11 siRNA and then treated with TGFβ as in ( A ), followed by co-culturing with HLFs for 48 h. HLFs were harvested and analyzed for ACTA2 and COL1A1 mRNA by qPCR. ( F ) RNA samples were isolated from A549 cells after TGFβ or PBS treatment and analyzed by qPCR for WNT3a, AXIN2 and WNT11 gene expression (fold-change from respective untreated control). ( G ) WNT11 mRNA expression was measured by qPCR in iHAEC2s after siRNA knockdown and TGFβ treatment. ( H ) Human lung fibroblast ACTA2 mRNA was measured following co-culture with control or Wnt11-siRNA-treated ihAEC2s with or without TGFβ. N = 3–4 in ( A – F ). p < * 0.05, ** 0.01, *** 0.001, **** 0.0001 between the indicated two groups by one-way ANOVA plus Tukey’s or Student’s t -test.

Article Snippet: Six-week-old female C57BL/6, Wnt11 fl/fl (B6;129-Wnt11tm.1Ser/J, Stock# 030051) and Col-Cre +/− (C57BL/6J-Tg[Col1α2-Cre-ER(T)], stock# 029567) mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Derivative Assay, Isolation, Transfection, Cell Culture, Expressing, Control, Gene Expression, Knockdown, Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Non-Canonical Wnt11 Signaling Regulates Pulmonary Fibrosis via Fibroblast and Alveolar Epithelial Type II Cell Crosstalk

doi: 10.3390/ijms27010351

Figure Lengend Snippet: Regulation of Wnt11 transcription and expression. ( A ) Four serial truncated human WNT11 promoter plasmid constructs (P1–P4) are illustrated. The purple box indicates the TCF/LEF binding element, while the green boxes are locations of putative Smad binding elements, with the red box also indicating a single CAGA sequence. ( B ) Four WNT11 promoter constructs were transfected into primary HLFs, respectively, and treated with Wnt3a (200 ng/mL) or CHIR 99021 (CHIR, 5 µM) for 24 h. WNT11 promoter activity was determined after normalization to respective Renilla luciferase activity. ( C ) HLFs were treated with TGFβ for 24 h and analyzed by ChIP assay for the binding of Smad3 to the WNT11 promoter using the primer sets bracketing R1, R2 and R3 regions. The results were expressed as fold-change from their own IgG controls. ( D ) HLFs transfected with each WNT11 promoter were treated with TGFβ (5 ng/mL) for 6 h; then, the activity was examined as described in ( B ). ( E ) CHIR- (5 µM) or DMSO-treated HLF lysates were analyzed by ChIP assay for β-catenin or TCF4 binding to the α-SMA promoter region as in ( B ). N = 3–4. Numbers above the bars indicate fold-change from unstimulated cells. N = 4–7. p < * 0.05, ** 0.01, *** 0.001, **** 0.0001 by one-way ANOVA with Tukey’s or Student’s t -test.

Article Snippet: Six-week-old female C57BL/6, Wnt11 fl/fl (B6;129-Wnt11tm.1Ser/J, Stock# 030051) and Col-Cre +/− (C57BL/6J-Tg[Col1α2-Cre-ER(T)], stock# 029567) mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Plasmid Preparation, Construct, Binding Assay, Sequencing, Transfection, Activity Assay, Luciferase

Journal: International Journal of Molecular Sciences

Article Title: Non-Canonical Wnt11 Signaling Regulates Pulmonary Fibrosis via Fibroblast and Alveolar Epithelial Type II Cell Crosstalk

doi: 10.3390/ijms27010351

Figure Lengend Snippet: Wnt signaling regulates myofibroblast differentiation. Canonical Wnt signaling is activated when Wnt3a binds to the LRP and Frizzled receptors, triggering an intracellular signaling cascade. Assembly of the destruction complex, comprising GSK-3β, CKIα, Axin and APC, promotes β-catenin phosphorylation and degradation; however, Wnt activation inhibits this process, allowing β-catenin to accumulate and translocate into the nucleus. Nuclear β-catenin then associates with TCF/LEF transcription factors to induce expression of target genes such as Wnt11. Wnt11 can be secreted by fibroblasts and activate non-canonical Wnt signaling by binding to Frizzled receptors and the LRP complex, leading to activation of the JNK/c-Jun pathway and subsequent transcription of Acta2. During bleomycin-induced lung injury, increased TGF-β further stimulates alveolar type II epithelial cells to secrete Wnt11, thereby enhancing paracrine activation of the non-canonical pathway. In addition, TGF-β upregulates Wnt11 transcription through Smad3-dependent mechanisms. Together, canonical and non-canonical Wnt signaling coordinate fibroblast-to-myofibroblast differentiation through distinct yet complementary pathways. Figure generated with BioRender. Created in BioRender. Gonzalez De Los Santos, F. (2025). https://BioRender.com/dvqk9o9 .

Article Snippet: Six-week-old female C57BL/6, Wnt11 fl/fl (B6;129-Wnt11tm.1Ser/J, Stock# 030051) and Col-Cre +/− (C57BL/6J-Tg[Col1α2-Cre-ER(T)], stock# 029567) mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Phospho-proteomics, Activation Assay, Expressing, Binding Assay, Generated

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cecal growth factors promote enteric neurosphere formation and hindgut colonization in the avian model

doi: 10.3389/fcell.2025.1681844

Figure Lengend Snippet: Isolation, characterization and differentiation of chicken enteric neurospheres. Schematic figure of avian enteric neurosphere generation (A) . Brightfield images show examples of neurospheres generated from E8 ENSCs that emerged 1, 3, and 7 days in GDNF-containing medium (B–D) . Representative immunofluorescence images of 7-day-old neurospheres generated from E8 chicken intestine (E–G) . Samples were cultured under three different conditions: Exp.1: control medium ( (E) , CTRL), Exp. 2: GDNF containing cell culture media ( (F) , GDNF), and Exp.3: GWEN ( (G) , ceca-derived mesenchymal growth factors: GDNF, WNT11, ET-3, Noggin) supplemented media. Immunofluorescence of cross-sections confirms the presence of neurons expressing TUJ1, and neural crest cells expressing PHOX2B. Compared to CTRL and GDNF-only conditions, GWEN treatment resulted in a robust increase in TUJ1+/PHOX2B+ cell aggregates on the surface of the neurosphere and single Phox2b+ cells in the center of the aggregates. The inset shows a magnified view of a PHOX2B+/TUJ1+ aggregate, indicating enhanced differentiation of neural crest derivatives (E–G) . Scale bar = 100 µm. CTRL, control; NS, neurosphere.

Article Snippet: To assess the effect of ceca-specific mesenchymal factors on enteric neural stem cells, the culture medium was further supplemented with 10 ng/mL glial cell line-derived neurotrophic factor (GDNF; R&D Systems, 212-GD-010), 200 ng/mL WNT11 (Origene, TP761904), 250 ng/mL endothelin-3 (ET-3; R&D Systems, 1,162/100U), and 100 ng/mL Noggin (R&D Systems, 6997-NG-025).

Techniques: Isolation, Generated, Immunofluorescence, Cell Culture, Control, Derivative Assay, Expressing