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: Animal Nutrition

Article Title: Methionine supplementation regulates eggshell quality and uterine transcriptome in late-stage broiler breeders

doi: 10.1016/j.aninu.2024.04.026

Figure Lengend Snippet: Regulatory effects of dietary Met on epithelial cell proliferation in the shell gland of broiler breeders in the late laying period. (A) Immunofluorescence identification of Wnt family member 11 ( WNT11 ) expression in the shell gland epithelial cells of the CON and Met groups. (B) Comparative analysis of the average positive staining area percentage for W NT 11 . (C) Relative mRNA expression of W NT 11 . These data are expressed as mean ± SEM for a sample size of n = 6. Double asterisks indicate highly significant differences ( P < 0.01).

Article Snippet: Sections were incubated with 5% bovine serum albumin, followed by overnight incubation at 4 °C with WNT family member 11 (WNT11) antibody (ImmunoWay Biotechnology, USA), with normal goat serum serving as the negative control.

Techniques: Immunofluorescence, Expressing, Staining

Journal: Animal Nutrition

Article Title: Methionine supplementation regulates eggshell quality and uterine transcriptome in late-stage broiler breeders

doi: 10.1016/j.aninu.2024.04.026

Figure Lengend Snippet: Interaction relationships between biomarkers related to eggshell biomineralization and selected differentially expressed genes (DEGs). (A) Interaction network between biomarkers and selected DEGs, where the lines represent existing interaction relationships. The greater the number of lines, the stronger and more numerous the interactions between the genes. (B) Correlation analysis between biomarkers and selected DEGs. The intensity of the color indicates the strength of the correlation, n = 6. FZD 1 = Frizzled receptor 1; CAMK1D = calcium/calmodulin-dependent protein kinase 1D; ATP2B2 = ATPase plasma membrane Ca 2+ transporting 2; EREG = epiregulin; AREG = amphiregulin; VEGFA = vascular endothelial growth factor A; FGF1 = fibroblast growth factor 1; ABCC9 = ATP binding cassette subfamily C member 9; SLC26A9 = solute carrier family 26 member 9; OC-116 = ovocleidin-116; OPN 3 = osteopontin 3; WNK1 = WNK lysine deficient protein kinase 1; CALB1 = calbindin 1; ITM2C = integral membrane protein 2C; MEPE = matrix extracellular phosphoglycoprotein; KCNJ3 = ATP-Sensitive inward rectifier potassium channel 3; RARRES1 = retinoic acid receptor responder 1; WNT11 = wnt family member 11; PSTA1 = prostate specific transcript 1; EPHA2 = EPH receptor A2; ULK1 = Unc-51 like autophagy activating kinase 1; NR4A1 = nuclear receptor subfamily 4 group a member 1; DUSP8 = dual specificity phosphatase 8; OXTR = oxytocin receptor; DAAM1 = dishevelled associated activator of morphogenesis 1; FOS = Fos proto-oncogene, AP-1 transcription factor subunit; SGK1 = serum/glucocorticoid regulated kinase 1.

Article Snippet: Sections were incubated with 5% bovine serum albumin, followed by overnight incubation at 4 °C with WNT family member 11 (WNT11) antibody (ImmunoWay Biotechnology, USA), with normal goat serum serving as the negative control.

Techniques: Clinical Proteomics, Membrane, Binding Assay