rabbit polyclonal antibodies against wnt5a  (Bioss)

Journal: Nature Communications

Article Title: Decreased scleral Wnt5a hi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

doi: 10.1038/s41467-025-67246-x

Figure Lengend Snippet: a Distribution of fibroblast subtypes expressed as a difference between FD-treated and FD-fellow sclera (two-sided chi-square test with Bonferroni correction for multiple testing). The X-axis indicates the subtypes corresponding to the uniform manifold approximation and projection plot. The Y-axis shows the log-transformed P -value obtained by the chi-square test between FD-treated and FD-fellow eyes’ scleral fibroblast subtypes. Positive values indicate a decrease, whereas negative values indicate an increase in the FD-treated eyes when compared with the FD-fellow eyes. b Heat map of the ECM outgoing (left) and incoming (right) of cell-cell interaction in all fibroblast subtypes. c Heat map of the outgoing (left) and incoming (right) cell-cell interaction signals of all fibroblast subtypes. The row indicates the relative strength of each interaction pathway. The column indicates the total interaction strength of each fibroblast subtype. d Ligand-receptor pairs of the ncWNT signalling pathway of all fibroblast subtypes. The relative contribution of each ligand-receptor pair of the ncWNT signalling pathway among all scleral fibroblast subtypes. e Expression profile of Wnt5a of ncWNT signalling in each subtype of fibroblast. f Expression profile of Wnt11 of ncWNT signalling in each fibroblast subtype. g Wnt5a mRNA level in FD-48h treated eyes and FD-48h fellow eyes (two-sided Wilcoxon signed-rank test). h Information flow of each cell–cell interaction pathway in FD treatment for 24 h or 48 h. FD form deprivation. Data are expressed as mean ± SEM.

Article Snippet: The sections were blocked with 6% normal donkey serum, 1% BSA, and 0.2% Triton X-100 in 0.1 M PBS for 2 h at room temperature, then incubated with rabbit polyclonal antibodies against WNT5A (1:100, #bs-1948R, Bioss, Woburn, MA, USA) and SPARC (1:300, #15274-1-AP, Proteintech, Rosemont, IL, USA) for 24 h at 4 °C.

Techniques: Transformation Assay, Expressing

Journal: Nature Communications

Article Title: Decreased scleral Wnt5a hi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

doi: 10.1038/s41467-025-67246-x

Figure Lengend Snippet: a Locations of regionally differentiated sclera: iPPS, oPPS, and peripheral sclera. b Alteration of the number and percentage of Wnt5a + A2m + cells and Wnt5a + puncta in the temporal and nasal iPPS, oPPS, and peripheral sclera in normal murine eyes at age 3 weeks. n = 18 micrographs; Friedman test with Dunn’s post hoc test (two-sided) or RM-ANOVA with Bonferroni’s post hoc test (two-sided). c , d The numbers and percentages of Wnt5a + A2m + cells and Wnt5a + puncta in the temporal/nasal iPPS, oPPS, and peripheral sclera in NC eyes, FD-F eyes, and FD-T eyes. n = 9 micrographs for NC eyes; for FD eyes, n = 12 micrographs in FD-24h and n = 9 micrographs in FD-48h, one-way ANOVA with Bonferroni’s post hoc test or Kruskal-Wallis non-parametric test with Dunn’s post hoc test (two-sided). e The Wnt5a mRNA level in NC eyes, FD-F eyes, and FD-T eyes treated by FD-48 h or FD-2W. For NC eyes, n = 10 in FD-48 h and n = 18 in FD-2W; for FD-F eyes and FD-T eyes, n = 9; one-way ANOVA with Bonferroni’s post hoc test. WNT5A protein immunofluorescence intensity ( f ) and distribution ( g ) in the temporal/nasal iPPS, oPPS, and peripheral sclera in normal murine eyes. n = 11, Friedman tests with Dunn’s post hoc tests (two-sided). WNT5A protein immunofluorescent intensity ( h ) and change ( i ) in the temporal/nasal iPPS, oPPS, and peripheral sclera in NC eyes, FD-F eyes, and FD-T eyes. For NC eyes, n = 5 in FD-48 h and n = 6 in FD-2W; for FD-F eyes and FD-T eyes, n = 5, one-way ANOVA with Bonferroni’s post hoc test or Kruskal–Wallis non-parametric test with Dunn’s post hoc test (two-sided). Western blots ( j ) and densitometric quantification ( k ) of scleral WNT5A protein in FDM mice. n = 6, one-way ANOVA with Bonferroni’s post hoc test (two-sided). red: Wnt5a ; green: A2m ; blue: 4’, 6-diamidino-2-phenylindole (DAPI), iPPS the inner peripapillary sclera, oPPS the outer peripapillary sclera, peripheral the peripheral sclera. NC normal control eyes, FD-F untreated contralateral eyes of form deprivation mice, FD-T form-deprived eyes. Data are expressed as mean ± SEM.

Article Snippet: The sections were blocked with 6% normal donkey serum, 1% BSA, and 0.2% Triton X-100 in 0.1 M PBS for 2 h at room temperature, then incubated with rabbit polyclonal antibodies against WNT5A (1:100, #bs-1948R, Bioss, Woburn, MA, USA) and SPARC (1:300, #15274-1-AP, Proteintech, Rosemont, IL, USA) for 24 h at 4 °C.

Techniques: Immunofluorescence, Western Blot, Control

Journal: Nature Communications

Article Title: Decreased scleral Wnt5a hi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

doi: 10.1038/s41467-025-67246-x

Figure Lengend Snippet: a sh Wnt5a -AAV injection and refraction and ocular biometrics measurements were carried out at different time points. b AAV infection in the whole murine sclera after sh Wnt5a sub-Tenon’s injection. c The scleral Wnt5a mRNA level declined by sh Wnt5a -AAV injection. n = 6 for sh Scramble -AAV injected group and n = 7 for sh Wnt5a -AAV injected group, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 post hoc test (two-sided). d , e WNT5A protein levels were significantly downregulated by sh Wnt5a -AAV injection. n = 7 mice per group, Friedman test with Dunn’s post hoc test (two-sided). f The refraction of sh Wnt5a -AAV injected eyes underwent a greater myopic shift than did the shScramble-AAV injected mice. n = 15 for the shScramble-AAV injected group and n = 13 for the sh Wnt5a -AAV injected group, two-way repeated measures ANOVA with Bonferroni’s post hoc test. g Compared with shScramble-AAV infection, sh Wnt5a -AAV infection caused a significant increase in axial length. n = 15 for the shScramble-AAV injected group and n = 13 for the sh Wnt5a -AAV injected group, two-way repeated measures ANOVA with Bonferroni’s post hoc test. Col1a1 and Acta2 mRNA ( h , for Col1a1 , n = 6 for shScramble-AAV injected group and n = 10 for sh Wnt5a -AAV injected group, Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 post hoc test, two-sided; for Acta2 , n = 7 for shScramble-AAV injected group and n = 9 for sh Wnt5a -AAV injected group, one-way ANOVA with Bonferroni’s post hoc test, two-sided) and protein ( i , j, n = 7 for COL1A1 and n = 3 for α-SMA, RM-ANOVA with Bonferroni’s post hoc test (two-sided) for COL1A1 and Friedman test with Dunn’s post hoc test (two-sided) for α-SMA) level in shScramble-AAV, sh Wnt5a -AAV injected mice. k scleral fibril ultrastructure and scleral fibril diameter of the fellow eyes and the treated eyes in shScramble-AAV injected mice and sh Wnt5a -AAV injected mice ( n = 3 mice per group). Data are expressed as mean ± SEM.

Article Snippet: The sections were blocked with 6% normal donkey serum, 1% BSA, and 0.2% Triton X-100 in 0.1 M PBS for 2 h at room temperature, then incubated with rabbit polyclonal antibodies against WNT5A (1:100, #bs-1948R, Bioss, Woburn, MA, USA) and SPARC (1:300, #15274-1-AP, Proteintech, Rosemont, IL, USA) for 24 h at 4 °C.

Techniques: Injection, Infection

Journal: Nature Communications

Article Title: Decreased scleral Wnt5a hi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

doi: 10.1038/s41467-025-67246-x

Figure Lengend Snippet: Six weeks after sub-Tenon injection of OE Wnt5a -AAV, Wnt5a was overexpressed in the sclera at mRNA ( a , n = 5 for vector -AAV injected group and n = 9 for OE Wnt5a -AAV injected group, Kruskal–Wallis non-parametric test with Dunn’s post hoc test) and protein levels ( b , c, n = 3, RM-ANOVA with Bonferroni’s post hoc test, two-sided). d Schematic schedule of FD treatment and Wnt5a overexpression with AAV injection. In the FDM mouse model, OE Wnt5a -AAV injected + FDM mice developed less myopic shift than did the vector-AAV injected mice ( e ). In comparison with the vector-AAV injected eyes, OE Wnt5a -AAV injected + FD eyes exhibited a significant difference for axial length ( f, n = 13 for vector-AAV injected group and n = 16 for OE Wnt5a -AAV injected group, two-way repeated measures ANOVA with Bonferroni’s post hoc tests, two-sided, for refraction and for axial length). g , h Wnt5a overexpression can partially rescue the COL1A1 protein level, which is downregulated by FD. n = 7, ordinary two-way ANOVA with Bonferroni’s post hoc test (two-sided). In mice undergoing normal refractive development ( i ), sub-Tenon’s injection of OE Wnt5a -AAV did not affect refraction ( j ) or axial length ( k ). n = 11 for the vector-AAV injected group and n = 15 for the OE Wnt5a -AAV injected group, two-way repeated measures ANOVA with Bonferroni’s post hoc test (two-sided). FD form deprivation; FDM form deprivation myopia. Data are expressed as mean ± SEM.

Article Snippet: The sections were blocked with 6% normal donkey serum, 1% BSA, and 0.2% Triton X-100 in 0.1 M PBS for 2 h at room temperature, then incubated with rabbit polyclonal antibodies against WNT5A (1:100, #bs-1948R, Bioss, Woburn, MA, USA) and SPARC (1:300, #15274-1-AP, Proteintech, Rosemont, IL, USA) for 24 h at 4 °C.

Techniques: Injection, Plasmid Preparation, Over Expression, Comparison

Journal: Nature Communications

Article Title: Decreased scleral Wnt5a hi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

doi: 10.1038/s41467-025-67246-x

Figure Lengend Snippet: a , b Gene ontology (GO) and Reactome pathway analysis of differentially expressed genes (DEGs) from scleral bulk RNA-seq after sub-Tenon’s injection of sh Wnt5a -AAV ( n = 2 each group, one-sided hyper-geometric test with Benjamini-Hochberg correction for multiple testing). c DEGs heatmap of the ECM pathway from scleral bulk RNA-seq analysis. d Reverse Transcription quantitative Polymerase Chain Reaction validation of the DEGs ( Sparc , Adamts2 , Pcolce2 , Pcolce , Bmp1 , Itga2 ) from bulk RNA-seq analysis. For Sparc, n = 4 for shScramble-AAV injected group and n = 6 for sh Wnt5a -AAV injected group; for Adamts2 , Pcolce2 and Bmp1, n = 5 for both shScramble-AAV injected group and sh Wnt5a -AAV injected group; for Pcolce, n = 3 for shScramble-AAV injected group and n = 5 for sh Wnt5a -AAV injected group; for Itga2 , n = 4 for shScramble-AAV injected group and n = 5 for sh Wnt5a -AAV injected group. One-way ANOVA with Bonferroni’s post hoc test (two-sided) for Sparc, Adamts2 , and Pcolce , Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 post hoc tests (two-sided) for Pcolce2, Bmp1 , and Itga2 . e , f Network of the 20 hub genes and functional enrichment (one-sided hyper-geometric test with Benjamini–Hochberg correction for multiple testing) of the Wnt5a assigned gene module. g Western blots and densitometric quantification of scleral WNT5A and SAPRC proteins; representative blots are shown for sh Wnt5a -AAV injected mice. n = 5, Friedman tests with Dunn’s post hoc test (two-sided) for WNT5A; RM-ANOVA with Bonferroni’s post hoc test (two-sided) for SAPRC. h , i Immunofluorescence intensity and quantification of SPARC protein expression in the temporal/nasal iPPS, oPPS, and peripheral sclera of NC eyes. In FD-48h, n = 6 mic per group; in FD-2W, n = 3 for NC eyes and n = 4 for both FD-F and FD-T eyes. One-way ANOVA with Bonferroni’s post hoc test (two-sided), Brown-Forsythe and Welch ANOVA tests with Dunnett’s T3 post hoc test (two-sided) or Kruskal–Wallis non-parametric test with Dunn’s post hoc test (two-sided) were used. Red: SPARC; blue: 4’, 6-diamidino-2-phenylindole (DAPI); iPPS the inner peripapillary sclera, oPPS the outer peripapillary sclera, peripheral, the peripheral sclera. NC normal control eyes, FD-F untreated contralateral eyes of form deprivation mice, FD-T form deprivation treated eyes. Data are expressed as mean ± SEM.

Article Snippet: The sections were blocked with 6% normal donkey serum, 1% BSA, and 0.2% Triton X-100 in 0.1 M PBS for 2 h at room temperature, then incubated with rabbit polyclonal antibodies against WNT5A (1:100, #bs-1948R, Bioss, Woburn, MA, USA) and SPARC (1:300, #15274-1-AP, Proteintech, Rosemont, IL, USA) for 24 h at 4 °C.

Techniques: RNA Sequencing, Injection, Reverse Transcription, Real-time Polymerase Chain Reaction, Biomarker Discovery, Functional Assay, Western Blot, Immunofluorescence, Expressing, Control

Journal: Nature Communications

Article Title: Decreased scleral Wnt5a hi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

doi: 10.1038/s41467-025-67246-x

Figure Lengend Snippet: Abnormal visual stimulation propagating via the retina-choroid-sclera signalling cascade reduces Wnt5a hi fibroblast numbers and/or activity, and thus Wnt5a secretion. Subsequently, the expression of genes for ECM molecules such as Col1a1 , Sparc , Pcolce , Pcelce2 , Adamts2 , Bmp1 , and Igta2 is downregulated via the noncanonical Wnt signalling pathway (Wnt5a-Ca 2+ -CaMKII). This response may disrupt ECM structural organisation, decrease collagen I level, and reduce the collagen fibril diameter, finally resulting in myopia progression due to increased distension of the posterior sclera under constant intraocular pressure.

Article Snippet: The sections were blocked with 6% normal donkey serum, 1% BSA, and 0.2% Triton X-100 in 0.1 M PBS for 2 h at room temperature, then incubated with rabbit polyclonal antibodies against WNT5A (1:100, #bs-1948R, Bioss, Woburn, MA, USA) and SPARC (1:300, #15274-1-AP, Proteintech, Rosemont, IL, USA) for 24 h at 4 °C.

Techniques: Activity Assay, Expressing

Journal: Pharmaceutical biology

Article Title: Yiguanjian decoction inhibits macrophage M1 polarization and attenuates hepatic fibrosis induced by CCl 4 /2-AAF.

doi: 10.1080/13880209.2021.1961820

Figure Lengend Snippet: Figure 3. YGJ inhibits the activation of M1 macrophages and non-canonical Wnt signalling pathways and inhibits the differentiation of hepatic progenitor cells into myofibroblasts in vivo. (a) STAT1, IRF3, IRF5, IRF8, and SOCS3 protein bands were depicted in the immunoblot images, and (b) the densitometric quantification of the protein bands presented as a histogram (n ¼ 5 per group). (c) The mRNA expressions of Wnt-4, -5 A, -5B, FZD-2, -3, and -6 in liver were measured by RT-PCR and nor- malized to GAPDH mRNA (n ¼ 5 per group). (d) Wnt5A, Wnt5B, and FZD2 protein bands were depicted in the immunoblot images, and (e) the densitometric quantifi- cation of the protein bands presented as a histogram (n ¼ 5 per group). (f) OV6, SOX9, EpCAM, CK19, and Hep mRNA expressions were measured by RT-PCR and normalized to GAPDH mRNA (n ¼ 5 per group). (g) EpCAM protein bands were depicted in the immunoblot images, and (h) the densitometric quantification of the protein bands presented as a histogram (n ¼ 5 per group). p < 0.05 and p < 0.01. N: untreated group (control); 2-AAF/CCl4: 2-acetylaminofluorene/carbon tetra- chloride-treated group; YGJ: 2-AAF/CCl4 þ Yiguanjian decoction-treated group; SORA: 2-AAF/CCl4 þ sorafenib-treated group.

Article Snippet: Rabbit polyclonal antibodies against Wnt5A and Wnt5B were purchased from Signalway Antibody LLC (MD, USA).

Techniques: Activation Assay, In Vivo, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Pharmaceutical Biology

Article Title: Yiguanjian decoction inhibits macrophage M1 polarization and attenuates hepatic fibrosis induced by CCl 4 /2-AAF

doi: 10.1080/13880209.2021.1961820

Figure Lengend Snippet: YGJ inhibits the activation of M1 macrophages and non-canonical Wnt signalling pathways and inhibits the differentiation of hepatic progenitor cells into myofibroblasts in vivo . (a) STAT1, IRF3, IRF5, IRF8, and SOCS3 protein bands were depicted in the immunoblot images, and (b) the densitometric quantification of the protein bands presented as a histogram ( n = 5 per group). (c) The mRNA expressions of Wnt-4, -5 A, -5B, FZD-2, -3, and -6 in liver were measured by RT-PCR and normalized to GAPDH mRNA ( n = 5 per group). (d) Wnt5A, Wnt5B, and FZD2 protein bands were depicted in the immunoblot images, and (e) the densitometric quantification of the protein bands presented as a histogram ( n = 5 per group). (f) OV6, SOX9, EpCAM, CK19, and Hep mRNA expressions were measured by RT-PCR and normalized to GAPDH mRNA ( n = 5 per group). (g) EpCAM protein bands were depicted in the immunoblot images, and (h) the densitometric quantification of the protein bands presented as a histogram ( n = 5 per group). * p < 0.05 and ** p < 0.01. N: untreated group (control); 2-AAF/CCl 4 : 2-acetylaminofluorene/carbon tetrachloride-treated group; YGJ: 2-AAF/CCl 4 + Yiguanjian decoction-treated group; SORA: 2-AAF/CCl 4 + sorafenib-treated group.

Article Snippet: Rabbit polyclonal antibodies against Wnt5A and Wnt5B were purchased from Signalway Antibody LLC (MD, USA).

Techniques: Activation Assay, In Vivo, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control