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: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: The level of serum Wnt5a or Wnt11 was measured by ELISA analysis. A , B The level of serum Wnt5a or Wnt11 in hypertension group (HBP, n = 56) and control group (Ctrl, n = 33); C , D The level of serum Wnt5a or Wnt11 in SBP ≥ 140 mmhg group ( n = 22) and SBP < 140 mmhg group ( n = 34); E , F The level of serum Wnt5a or Wnt11 in DBP ≥ 90 mmhg group ( n = 16) and DBP < 90 mmhg group (DBP, n = 40); G , H The E/A ratio between the healthy control group (Ctrl) and the hypertension (HBP) group was detected by echocardiography, and the ratio was divided into E/A ≥ 1 ( n = 43) and E/A < 1 ( n = 46). The level of serum Wnt5a or Wnt11 in the two groups were analyzed. SBP systolic blood pressure, DBP diastolic blood pressure. All data were expressed as means ± SEM.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A The expressions of Wnt5a and Wnt11 were analyzed by western blot in the heart tissues of patients with dilated cardiomyopathy (DCM) or control (Ctrl) group, n = 4/group. B Western blot analysis of Wnt5a and Wnt11 expressions in heart tissues from TAC mice at different time-points (3 days, 1 week, 2 weeks, 4 weeks), n = 5/group. C , D Western blot analysis of Wnt5a and Wnt11 expression in cardiac myocytes (CMs), fibroblasts (CFs), and their supernatants at different time-points (6, 12, 24, 36, and 48 h) after mechanical stretch (MS), n = 6/group. All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Western Blot, Control, Expressing

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A Western blot analysis of Wnt5a and Wnt11 in heart tissues ( n = 6/group). B Cardiac fibrosis was analyzed by Masson staining in heart tissues ( n = 5/group). C Western blot analysis of Col-1, MMP2, MMP9, and TGF-β1 levels in heart tissues ( n = 5/group). Mice were injected with sh-Wnt5A /Wnt11-AAV9 (shWnt5a/11) or sh-Scramble-AAV9 (sh-NC) by tail vein, 2 weeks later, these mice were subjected to TAC or sham operation. After 4 weeks, further analysis was performed. D Western blot analysis of serum Wnt5a or Wnt11 level at 4 weeks after sham or TAC operation, and these mice were infused recombinant rat Wnt5a (rWnt5a) and Wnt11 (rWnt11), or vehicle (as control) by minipump for 2 weeks from the sham or TAC operation ( n = 6/group). E Cardiac fibrosis was analyzed by Masson staining in heart tissues ( n = 6/group). All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Western Blot, Staining, Injection, Recombinant, Control

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A , B Western analysis of Wnt5a or Wnt11 in stretched (MS) or control (Ctrl) cardiomyocytes pre-transfected sh-Scramble (sh-NC), sh-Wnt5a or sh-Wnt11 lentivirus, respectively. n = 5/group. C , D Western analysis of Col-1, MMP9, MMP2, and TGF-β1 in cardiac fibroblasts (CFs) treated with conditional medium from cardiomyocytes (CMs) in ( A ) or ( B ), respectively. n = 6/group Stretched (MS) or control (Ctrl) cardiomyocytes were pre-transfected with sh-Scramble (sh-NC), ()shWnt5a or shWnt11 lentivirus, respectively. The conditional medium of these cardiac myocytes was collected to treat fibroblasts for 24 h, and the expressions of Col-1, MMP9, MMP2, and TGF-β1 were detected by Western blot. All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Western Blot, Control, Transfection

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A , B Western analysis of Wnt5a, Wnt11, Col-1, α-SMA, MMP9, MMP2, and TGF-β1 in stretched (MS) or control (Ctrl) cardiac fibroblasts pre-transfected with sh-Wnt5a, sh-Wnt11 or sh-scramble (sh-NC) lentivirus, respectively. n = 6/group. All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Western Blot, Control, Transfection

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A The expression of Col-1, MMP9, MMP2, and TGF-β1 were detected by western blot analysis in cardiac fibroblasts stimulated with exogenous recombinant rat Wnt5a (rWnt5a) or Wnt11 (rWnt11) at different concentrations (0, 10, 50, 100 ng/ml) for 24 h. n = 6/group. B The levels of p-EGFR, p-ERK, and p-P38 were examined in cardiac fibroblasts stimulated with exogenous recombinant rat Wnt5a (rWnt5a) or Wnt11 (rWnt11) at different concentrations (0, 10, 50, 100 ng/ml) and different time-points (0, 5, 10, 30 min), n = 6/group. All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Expressing, Western Blot, Recombinant

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A , B The expression of EGFR, Col-1, α-SMA, MMP9, MMP2, and TGF- β1 were detected by western blot in cardiac fibroblasts. After transfection with si-RNA-EGFR (si-EGFR) or si-RNA-scramble (si-Scram) for 48 h, these fibroblasts were treated with recombinant rat Wnt5a (rWnt5a) or Wnt11 (rWnt11) for 24 h, respectively. The same volume of PBS was treated as a control. n = 6/group. C , D The levels of p-ERK, p-P38, FZD5, and EGFR were examined by western blot analysis in cardiac fibroblasts. These fibroblasts were transfected with si-RNA-EGFR (si-EGFR) or si-RNA-scramble (si-Scram) for 48 h and then treated with recombinant rat Wnt5a (rWnt5a) or Wnt11 (rWnt11), respectively, 30 min later, these cells were analyzed. n = 5/group. All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Expressing, Western Blot, Transfection, Recombinant, Control

Journal: Cell Death & Disease

Article Title: Cardiac Wnt5a and Wnt11 promote fibrosis by the crosstalk of FZD5 and EGFR signaling under pressure overload

doi: 10.1038/s41419-021-04152-2

Figure Lengend Snippet: A , B Western blot analysis of FZD5, Col-1, α-SMA, MMP9, MMP2, and TGF-β1 expression in cardiac fibroblasts. Si-RNA-FZD5 (si-FZD5) or si-RNA-scramble (si-Scram) was treated with fibroblasts for 48 h, then exogenous recombinant rat Wnt5a (rWnt5a) or Wnt11 (rWnt11) were added into cultured fibroblasts for 24 h for further analysis. n = 6/group. C , D Western blot analysis of p-EGFR, EGFR, p-ERK, p-P38, and FZD5 levels in cardiac fibroblasts. Si-RNA-FZD5 (si-FZD5) or si-RNA-scramble (si-Scram) was treated into fibroblasts for 48 h, then exogenous recombinant rat Wnt5a (rWnt5a) or Wnt11 (rWnt11) were added into cultured fibroblasts for 30 min for further analysis. n = 5/group. All data were shown as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In brief, blood samples, which we collected from patients, were centrifuged with 1000x g at 4 ° C for 20 min, and then the supernatant was collected for analysis of Wnt5a and Wnt11 level by ELISA Kit (CUSABIO, Wuhan, China) according to the manufacturer’s protocols.

Techniques: Western Blot, Expressing, Recombinant, Cell Culture

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 3 WNT11 is a novel ligand for ROR2 in humans. a RNA-Seq of MCF-7 pROR2 cells: Network of differentially expressed genes associated with non-canonical WNT signaling grouped according to their cellular localization. b MCF-7 cells were stimulated for 24 h with rWNT5A (100 ng/ml) and WNT11 expression was analyzed by western Blot. c+d Expression of the non-canonical WNT ligands was measured by qRT-PCR in MCF-7 (c) or the indicated ROR2-overexpressing human breast cancer cell lines (d) (mean ± SD, n = 3–9, *p < 0.05, p < 0.01, n.e. = not expressed). Expression values were calculated relative to the empty vector control cells. e Co-immunoprecipitation (Co-IP) of V5-WNT11 in MCF-7 pROR2 cells detects ROR2 by western blot. f Schematic representation of the ROR2 N-terminal deletion constructs. g Co-IP of V5-Wnt11 in MCF-7 expressing either pROR2-FL or pROR2-ΔΔ. h Cell invasion assays of MCF-7 expressing N-terminal ROR2 deletion constructs (mean ± SD, n = 3, *p < 0.01, **p = 0.0001, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: RNA Sequencing, Expressing, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Control, Immunoprecipitation, Co-Immunoprecipitation Assay, Construct, Comparison

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 4 WNT11 mediates the pro-tumoral effects of ROR2. a+b Invasion assay: MCF-7 pcDNA or pROR2 cells were treated with WNT inhibitors (a) or Porcupine inhibitors (b) (mean ± SD, n = 3, *p < 0.05, **p < 0.01, ***p < 0.0001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c Invasion assay of MCF-7 pROR2 cells stably expressing a non-sense control (ns ctl) or WNT11 (shWNT11) shRNA (mean ± SD, n = 3, *p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. d Invasion of MCF-7 pROR2 cells transfected with control (siCTL) or WNT11 siRNA (siWNT11) +/− rhWNT11 (100 ng/ml) was assessed in Boyden chambers (mean ± SD, n = 3, *p < 0.05, **p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. e Invasion assay of BT-474 pcDNA and pROR2 cells transfected with either siCTL or siWNT11 (mean ± SD, n = 3, *p < 0.05, **p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. f AFM of cell-cell-junctions in the indicated cell lines. g Immunofluorescence for the tight junction protein ZO-1. h ECIS measurements of MCF-7 cells (box: 25-75th percentile, line at median, *p < 0.0001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. i+j xCELLigence measurements of MCF-7 cells (mean ± SD). Shown is one representative example (i) and a corresponding Area Under the Curve (AUC) analysis for all three independent experiments (mean ± SD, *p < 0.01). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: Invasion Assay, Comparison, Stable Transfection, Expressing, Control, shRNA, Transfection, Immunofluorescence

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 5 WNT11 mediates ROR2 signaling. a+b Western blot: RHOA and ROCK2 in MCF-7 and BT-474 pROR2 cells treated with control siRNA (siCTL) or siRNA against WNT11 (siWNT11) (a) with corresponding densitometric quantification normalized on HSP90 expression (b) (mean ± SD, *p < 0.05, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c Levels of active RHOA-GTP in the indicated cells were assessed by ELISA (mean ± SD, *p < 0.05, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c MCF-7 pcDNA and pROR2 siCTL/siWNT11 cells were characterized by RPPA for phospho-proteins associated with the WNT signaling pathway (n = 3, *p < 0.05). d Schematic representation of the master regulator analysis of ROR2/WNT11 signaling. The illustration was created with BioRender.com

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: Western Blot, Control, Expressing, Comparison, Enzyme-linked Immunosorbent Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 6 ROR2/WNT11 are expressed in metastatic breast cancer and associated with poor survival. a Pathway (pw) enrichment for RNA-Seq data from 31 patients with brain metastases given for different WNT subpathways. Significance was calculated with a log rank test. b qRT-PCR: Expression of ROR2 and WNT11 in samples of human brain metastases (line at median). The upper numbers indicate the number of samples with positive signals out of all investigated samples. c Kaplan-Meier survival curves showing the OS of metastatic patients based on their averaged WNT11 and ROR2 expression. The separation high/low was computed based on an optimal cutoff using the maxstat method. Significance was calculated with a log rank test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: RNA Sequencing, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: A homozygous human WNT11 loss-of-function variant associated with laterality, heart and renal defects

doi: 10.1101/2024.11.14.623711

Figure Lengend Snippet: (A) Pedigree indicating clinical features and segregation of the identified WNT11 c.814delG variant amongst the three family members. (B) Chromatogram traces of Sanger sequencing results regarding the WNT11 variant segregation within the family, showing a homozygous deletion of c.814G in the affected child while both parents are heterozygous. In (A,B) , I.1 = father; I.2 = mother; II.1 = son. (C,D) Alphafold 2.1.2 protein structure predictions of human (WNT11) and Xenopus (Wnt11b) Wnt ligands, and comparison the WNT11 c.814delG variant to dominant-negative (dn)Wnt11b. (D) Magnification of C-termini of truncated ligands with annotation of the last amino acids. Red arrows indicate C-terminal end.

Article Snippet: Human WNT11 ( HWNT11-pCMV6-Entry ) was obtained from OriGene (#RC219688) (matching ENST00000322563.8, NM_004626.3, CCDS8242) and subcloned into pCS108 (using Cla1 and EcoR1 enzymes; NEB, #R0197 and #R3101) to serve as templates for in vitro mRNA synthesis using primers listed below.

Techniques: Variant Assay, Sequencing, Comparison, Dominant Negative Mutation

Journal: bioRxiv

Article Title: A homozygous human WNT11 loss-of-function variant associated with laterality, heart and renal defects

doi: 10.1101/2024.11.14.623711

Figure Lengend Snippet: (A-D) Overexpression of mRNAs encoding WNT11, Wnt11b, WNT11 c.814delG and dnWnt11b, and analysis of effects on (A,B) morphogenesis at st. 13 (posterior view) and 20 (dorso-anterior view), (C,D) and left-right axis patterning (lateral views) by WMISH at st. 30 - 32 against pitx2c . (A,D) Representative images. In (A) , neural folds (nf) and blastopores (bp) are indicated by yellow and magenta dashed lines, respectively. In (D) , filled green arrowheads indicate normal left-sided expression, open white arrowheads indicate normal right-sided lack of expression. Filled magenta arrowheads indicate ectopic right-sided expression, open magenta arrowheads indicate loss of left-sided expression. (B,C) , Quantification of results. n = number of embryos analyzed per condition. χ 2 -test, p > 0.05 = ns; p < 0.05 = *; p < 0.01 = **; p < 0.001 = ***.

Article Snippet: Human WNT11 ( HWNT11-pCMV6-Entry ) was obtained from OriGene (#RC219688) (matching ENST00000322563.8, NM_004626.3, CCDS8242) and subcloned into pCS108 (using Cla1 and EcoR1 enzymes; NEB, #R0197 and #R3101) to serve as templates for in vitro mRNA synthesis using primers listed below.

Techniques: Over Expression, Expressing

Journal: bioRxiv

Article Title: A homozygous human WNT11 loss-of-function variant associated with laterality, heart and renal defects

doi: 10.1101/2024.11.14.623711

Figure Lengend Snippet: (A) Schematic representation of constructs and their lengths. Human WNT11 constructs are shown in blue, Xenopus Wnt11b constructs are shown in green. Orange indicates changes in the sequence. (B-E) Overexpression of mRNAs encoding WNT11, WNT11 c.814delG , WNT11 D814-1062 , WNT11 D850-1062 , dnWnt11b and Wnt11b D811-1059 , and analysis of effects on (B,C) morphogenesis at st. 17 (dorso-anterior view), and (D,E) left-right axis patterning (lateral views) by WMISH at st. 30 - 32 against pitx2c . (B,E) Representative images. In (B) , neural folds (nf) are indicated by yellow dashed lines. In (E) , filled green arrowheads indicate normal left-sided expression, open white arrowheads indicate normal right-sided lack of expression. Filled magenta arrowheads indicate ectopic right-sided expression, open magenta arrowheads indicate loss of left-sided expression. (C,D) Quantification of results. n = number of embryos analyzed per condition. χ 2 -test, p > 0.05 = ns; p < 0.05 = *; p < 0.01 = **; p < 0.001 = ***.

Article Snippet: Human WNT11 ( HWNT11-pCMV6-Entry ) was obtained from OriGene (#RC219688) (matching ENST00000322563.8, NM_004626.3, CCDS8242) and subcloned into pCS108 (using Cla1 and EcoR1 enzymes; NEB, #R0197 and #R3101) to serve as templates for in vitro mRNA synthesis using primers listed below.

Techniques: Construct, Sequencing, Over Expression, Expressing

Journal: bioRxiv

Article Title: A homozygous human WNT11 loss-of-function variant associated with laterality, heart and renal defects

doi: 10.1101/2024.11.14.623711

Figure Lengend Snippet: (A,B) Overexpression of mRNAs encoding WNT11 c.814delG , WNT11 D814-1062 and WNT11 D850-1062 , and analysis of effects on Wnt/β-catenin signaling reporter (pbinLEF::dGFP) activity (green). (A) Representative images. (B) Quantification of results. n = number of embryos analyzed per condition. χ 2 -test, p > 0.05 = ns; p < 0.05 = *; p < 0.01 = **; p < 0.001 = ***. (C,D) Overexpression of mRNAs encoding WNT11, WNT11 c.814delG , WNT11 D814-1062 , WNT11 D850-1062 and Wnt11b, and analysis of (C) protein expression levels as well as (D) dimerization of ligands using Western blotting. α-WNT11 = anti-human WNT11 antibody; α-Tub. = anti-alpha-Tubulin antibody (loading control). Full membranes are shown in supplemental figure S2C and D.

Article Snippet: Human WNT11 ( HWNT11-pCMV6-Entry ) was obtained from OriGene (#RC219688) (matching ENST00000322563.8, NM_004626.3, CCDS8242) and subcloned into pCS108 (using Cla1 and EcoR1 enzymes; NEB, #R0197 and #R3101) to serve as templates for in vitro mRNA synthesis using primers listed below.

Techniques: Over Expression, Activity Assay, Expressing, Western Blot, Control