Journal: Nature Communications

Article Title: Norrin-induced Frizzled4 endocytosis and endo-lysosomal trafficking control retinal angiogenesis and barrier function

doi: 10.1038/ncomms16050

Figure Lengend Snippet: ( a ) WNT5A/PMA-induced FZD4 internalization in HeLa cells. WT DVL2 binds and co-localizes with FZD4 and does not form cytoplasmic aggregates. ( b ) DVL2 K466M is FZD4-binding deficient and thus forms cytoplasmic aggregates. In the DVL2 K466M expressing cells, WNT5A/PMA fails to induce FZD4 endocytosis. ( c ) DVL2 AHEL, which is impaired in AP2 binding, does not form cytoplasmic aggregates but also prevents WNT5A/PMA-induced internalization of FZD4. ( d – f ) NDP induces strong FZD4 endocytosis regardless of K466M or AHEL mutations in co-expressed DVL2. Scale bar, 10 μm.

Article Snippet: WNT5A CM was generated from L Wnt-5A cells (ATCC CRL-2814) stably expressing WNT5A.

Techniques: Binding Assay, Expressing

Journal: Nature Communications

Article Title: Norrin-induced Frizzled4 endocytosis and endo-lysosomal trafficking control retinal angiogenesis and barrier function

doi: 10.1038/ncomms16050

Figure Lengend Snippet: ( a – c ) FZD4 mutations that disrupt DVL2 binding fail to recruit DVL2 to the plasma membrane and block WNT5A/PMA-induced endocytosis. ( d – f ) Norrin-induced FZD4 endocytosis is not affected by FZD4 mutations that disrupt DVL2 binding. DVL2 was co-expressed in all a – f . Scale bar, 10 μm.

Article Snippet: WNT5A CM was generated from L Wnt-5A cells (ATCC CRL-2814) stably expressing WNT5A.

Techniques: Binding Assay, Clinical Proteomics, Membrane, Blocking Assay

Journal: Molecular medicine reports

Article Title: Effects of Wnt5a protein on proliferation and apoptosis in JAR choriocarcinoma cells.

doi: 10.3892/mmr.2010.383

Figure Lengend Snippet: Figure 2. Wnt5a expression in JAR cells, but not in primary CTB cells. (A) Analysis of Wnt5a expression in CTB and JAR cells by PCR electrophoresis. (B) Positive staining of Wnt5a in primary CTB cells. (C) Negative staining of Wnt5a in JAR cells. Bars, 50 µm.

Article Snippet: Jar cells were plated in 6-well plates 24 h before Brdu incorporation analysis. recombinant mouse Wnt5a (mWnt5a; 645-Wn; r&d Systems inc.) at concentrations of 0, 100, 200 and 400 ng/ml were added to the medium.

Techniques: Expressing, Electrophoresis, Staining, Negative Staining

Journal: Molecular medicine reports

Article Title: Effects of Wnt5a protein on proliferation and apoptosis in JAR choriocarcinoma cells.

doi: 10.3892/mmr.2010.383

Figure Lengend Snippet: Figure 4. Effect of mWnt5a proteins on the growth of JAR cells. Viable cells were counted from days 1-7. JAR cells without Wnt5a treatment were used as a control. The number of viable cells was higher in JAR-mWnt5a than in JAR cells from days 2-6 (P<0.05).

Article Snippet: Jar cells were plated in 6-well plates 24 h before Brdu incorporation analysis. recombinant mouse Wnt5a (mWnt5a; 645-Wn; r&d Systems inc.) at concentrations of 0, 100, 200 and 400 ng/ml were added to the medium.

Techniques: Control

Journal: Molecular medicine reports

Article Title: Effects of Wnt5a protein on proliferation and apoptosis in JAR choriocarcinoma cells.

doi: 10.3892/mmr.2010.383

Figure Lengend Snippet: Figure 5. Effect of mWnt5a proteins on the apoptosis of JAR cells. The percentage of Annexin V-positive cells in the serum-starvation culture was examined to evaluate the effect of Wnt5a on the apoptosis of JAR cells. (A) Representative results of flow cytometric analysis of JAR cells in 10% FBS, 0.5% FBS, and 0.5% FBS with mWnt5a. (B) Quantitative measurement of the percentage of Annexin V-positive cells. Analyses were performed three times in independent experiments, and 10,000 cells were counted in each case. *P<0.05.

Article Snippet: Jar cells were plated in 6-well plates 24 h before Brdu incorporation analysis. recombinant mouse Wnt5a (mWnt5a; 645-Wn; r&d Systems inc.) at concentrations of 0, 100, 200 and 400 ng/ml were added to the medium.

Techniques:

Journal: Experimental & Molecular Medicine

Article Title: Differential effects of type 1 diabetes mellitus and subsequent osteoblastic β-catenin activation on trabecular and cortical bone in a mouse model

doi: 10.1038/s12276-018-0186-y

Figure Lengend Snippet: a WNT16 and b WNT5a protein expressions were detected by immunohistochemistry in longitudinal sections of femoral trabecular and cortical bone. The quantitative results are shown. n = 9. Scale bars, 50 μm. c mRNA expression levels of WNT16 and WNT5a in trabecular and cortical bone of control mice. n = 8. d mRNA expression levels of WNT16 and WNT5a in trabecular bone and cortical bone in T1DM and T1-CA mice. n = 8. e mRNA expression levels and protein concentrations of WNT16 and WNT5a in primary osteoblasts from Catnblox(ex3) mice treated with high glucose plus Cre adenovirus or vehicle. n = 8. Ob osteoblast. Data are expressed as the mean ± SD. ** P < 0.01 versus trabecular bone ( a – c ), T1DM group ( d ) or Catnblox(ex3) osteoblasts ( e ) by an unpaired t -test. NS not significant, P > 0.05

Article Snippet: The primary antibodies used in the study included a rabbit monoclonal anti-β-catenin antibody (1:400; #8814; Cell Signaling), a rabbit polyclonal anti-Dickkopf-1 antibody (1:100; sc-25516; Santa Cruz), a mouse monoclonal anti-WNT16 antibody (1:100; sc-271897; Santa Cruz), and a rabbit polyclonal anti-WNT5a antibody (1:100; ab72583; Abcam).

Techniques: Immunohistochemistry, Expressing, Control

Journal:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

doi: 10.1111/j.1755-148X.2010.00792.x

Figure Lengend Snippet: (A) Total protein was isolated from melanoma cell lines of different metastatic potential and Klotho protein expression was measured by western blot analysis. β-Tubulin was used as a loading control. (B) Klotho mRNA expression by real-time PCR in G361, UACC903, and M93-047 melanoma cell lines normalized to 18S levels (n=4, error bars are standard deviation, *= p< 0.05, ** = p<0.005). (C) Percentages of lesions expressing Klotho, n= no of tumors per group. Prim= Primary melanoma, VM= Visceral metastases, LNM= Lymph node metastases. (D) Serial sections of primary melanoma stained for both Klotho and Wnt5A expression.

Article Snippet: After washing three more times, the plates were incubated with biotinylated Wnt5A antibody (R&D Systems, 1:500) overnight at 4oC.

Techniques: Isolation, Expressing, Western Blot, Control, Real-time Polymerase Chain Reaction, Standard Deviation, Staining

Journal:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

doi: 10.1111/j.1755-148X.2010.00792.x

Figure Lengend Snippet: The expression and localization of Klotho (red) was examined in melanoma cell lines expressing different levels of Wnt5A using immunofluorescent confocal microscopy. (A) The expression of Klotho was low in M93-047 and UACC903 cell lines and (B) was abrogated in G361 cells following treatment with recombinant Wnt5A for 16 hours. (C) Total protein lysates from G361 cells were analyzed by Western blot for the expression of Klotho, which was decreased following treatment with recombinant Wnt5A for 16 hrs. β-Tubulin was used as a loading control. (D) Analysis of Klotho mRNA expression measured by real-time PCR analysis shows that treatment of G361 cells with recombinant Wnt5A decreased Klotho expression (n=3, error bars are standard error of the mean, * = p<0.05). Transfection of Wnt5A-expressing M93-047 with Wnt5A siRNA for 48 hrs resulted in an increase in (E) Klotho mRNA expression (Klotho mRNA expression was normalized to 18S levels (n=3, error bars are standard error of the mean, ** = p<0.005)), as well as (F) Klotho protein expression.

Article Snippet: After washing three more times, the plates were incubated with biotinylated Wnt5A antibody (R&D Systems, 1:500) overnight at 4oC.

Techniques: Expressing, Confocal Microscopy, Recombinant, Western Blot, Control, Real-time Polymerase Chain Reaction, Transfection

Journal:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

doi: 10.1111/j.1755-148X.2010.00792.x

Figure Lengend Snippet: Total protein lysates from UACC903 and/or M93-047 cell lines were used to analyze Wnt5A and MART-1 protein expressions following treatment with recombinant Klotho for 16 hrs. (A) Wnt5A expression was decreased in both cell lines following rKlotho treatment. (B) MART-1 protein expression was increased in UACC903 following rKlotho treatment. β-Tubulin was used as a loading control. (C) Real-time PCR analysis of Wnt5A mRNA in M93-047 cells. Wnt5A expression was not significantly decreased in the presence of rKlotho. mRNA expression for Klotho was normalized to 18S levels (n=3, error bars are standard error of the mean). (D) Cells were treated with rKlotho for 16h, and the medium of the cells was examined for Wnt5A release. In the presence of Klotho, Wnt5A accumulates in the medium (upper panel), which correlates with a decrease in Wnt5A expression in total protein lysate (bottom panel). (E) Following appropriate treatment of cells, culture medium was collected and analyzed for sialidase activity as measured by sialic acid release. (Each sample was assayed in triplicate, n=3, error bars are standard error of the mean, * = p<0.05). (F) M93047 cells were treated for 16 hrs with recombinant Klotho in the presence or absence of DANA and total protein lysates were used to analyze Wnt5A protein expression. In the presence of DANA, the Klotho-induced decrease in Wnt5A expression is no longer observed.

Article Snippet: After washing three more times, the plates were incubated with biotinylated Wnt5A antibody (R&D Systems, 1:500) overnight at 4oC.

Techniques: Recombinant, Expressing, Control, Real-time Polymerase Chain Reaction, Activity Assay

Journal:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

doi: 10.1111/j.1755-148X.2010.00792.x

Figure Lengend Snippet: The expression and localization of Syndecan-1 (red) and Wnt5A (green) were examined in M93-047 cells using immunofluorescent confocal microscopy. (A) Treatment of M93-047 cells with recombinant Klotho for 16hrs shows that the decrease in Wnt5A expression in the presence of Klotho correlates with a decrease in the expression of Syndecan-1. (B) Co-localization of Wnt5A and syndecan-1 is affected as early as 5 mins after adding recombinant Klotho. (C) A solid phase binding assay demonstrates that in the presence of rKlotho, the binding of rWnt5A to rSyndecan is inhibited in a dose-dependent manner. (D) In the presence of DANA (50nM), treatment with recombinant klotho for 16 hrs no longer affects Wnt5A and syndecan expression levels as well as their co-localization.

Article Snippet: After washing three more times, the plates were incubated with biotinylated Wnt5A antibody (R&D Systems, 1:500) overnight at 4oC.

Techniques: Expressing, Confocal Microscopy, Recombinant, Binding Assay

Journal:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

doi: 10.1111/j.1755-148X.2010.00792.x

Figure Lengend Snippet: (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p<0.05, **=p<0.005. (B) Calpain-1 protein expression was measured by Western blot analysis and was decreased in the presence of Klotho. (C) Total protein lysates from M93-047 cells were analyzed by Western blot for the presence of Filamin A cleavage. The N-terminal Filamin A cleavage fragment was identified as a 190 kDa band, and cleavage was decreased in the presence of Klotho. β-Tubulin was used as a loading control. (D) Expression of the N-terminal Filamin A cleavage fragment was increased in G361 cells transfected with Klotho SiRNA (E) The effect of excess rWnt5A on the Klotho-mediated expression and localization of filamin was examined in M93-047 cells. (F) M2 cells were transfected with either wildtype (WT) or Calpain resistant (CR) Filamin A (FLNA) treated with recombinant Klotho and /or recombinant Wnt5A for 16 hrs. (G) Total protein lysates were analyzed for the expression of N-terminal Filamin A cleavage and, (H) the invasive potential of the cells was measured using a matrigel invasion assay as described in (A). Error bars are standard error of the mean, * = p<0.05, **=p<0.005.

Article Snippet: After washing three more times, the plates were incubated with biotinylated Wnt5A antibody (R&D Systems, 1:500) overnight at 4oC.

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

Journal: Molecular Brain

Article Title: NMDA receptor activation stimulates transcription-independent rapid wnt5a protein synthesis via the MAPK signaling pathway

doi: 10.1186/1756-6606-5-1

Figure Lengend Snippet: NMDAR activation rapidly increases Wnt5a in cortical cultures . A. Cellular localization of Wnt5a in neurons. Shown are confocal images of primary cortical neurons after double-fluorescent immunostaining with anti-Wnt5a (red) and anti-synapsin I (green) antibodies. The nucleus was stained by DAPI (blue). B. MSG or NMDA stimulation increased Wnt5a protein. Primary cortical neurons (10 DIV) were treated with 10 μΜ MSG or 50 μΜ NMDA for 15 min. Wnt5a protein was detected by Western blotting. Data in the summary graphs (mean ± SEM) were from three independent experiments (*, p < 0.05; **, p < 0.01; One-way ANOVA). C. NMDA receptor-regulated Wnt5a increase. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 100 μΜ DAP5 for 30 min, then incubated with 50 μΜ NMDA for 15 min. Wnt5a and p-P70S6K(included as a marker for translation activation) were detected by Western blotting summarized in the graph (n = 3; *, p < 0.05; **, p < 0.01; One-way ANOVA). D. Dynamic expression of Wnt5a protein after NMDA stimulation. Primary cortical neurons (10 DIV) were treated with 50 μΜ NMDA for 0, 5, 15, 30 and 60 min followed by Western blotting analysis of Wnt5a (n = 3; *, P < 0.05; **, p < 0.01; One-way ANOVA). E. NMDA-induced Wnt5a protein secretion. Primary cortical neurons (10 DIV) were treated with 50 μΜ NMDA for 0, 2, 4, 8, 16 and 32 min, and Wnt5a protein in the media was concentrated and detected on immunoblots.

Article Snippet: Anti-Wnt5a antibody was purchased from R&D Systems (AF654); anti-p-P70S6K (Thr389) antibody from Cell Signaling Technology (#9206); anti-GAPDH antibody from Santa Cruz (SC-32233); anti-Synapsin I from Millipore (AB1543P); and FITC-conjugated donkey anti-rabbit secondary antibody (711-097-020) and Rhodamine-conjugated rabbit anti-goat secondary antibody (305-297-003) from Jackson.

Techniques: Activation Assay, Immunostaining, Staining, Western Blot, Control, Incubation, Marker, Expressing

Journal: Molecular Brain

Article Title: NMDA receptor activation stimulates transcription-independent rapid wnt5a protein synthesis via the MAPK signaling pathway

doi: 10.1186/1756-6606-5-1

Figure Lengend Snippet: NMDAR-elicited Wnt5a increase requires translation but not transcription . A. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 20 μΜ anisomycin for 30 min, and then incubated with 50 μΜ NMDA for 15 min, followed by Wnt5a immunoblotting. The Graph is a summary of three independent experiments (**, p < 0.01; One-way ANOVA). B. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 20 μΜ actinomycin D for 30 min, followed by addition of 50 μΜ NMDA for 15 min. Wnt5a protein was detected by Western blotting and quantified. The Graph is a summary of four independent experiments (*, p < 0.05; **, p < 0.01; One-way ANOVA). C. Primary cortical neurons (10 DIV) were treated with vehicle (Control) or 50 μΜ NMDA for 15 min. Wnt5a mRNA was quantified by Real-time RT-PCR (qPCR). The summary graph is from three independent experiments (40 cycles, CT values: 25.1 ± 0.5/control vs. 25.6 ± 0.3/NMDA; p > 0.05; two-tailed Student's tests). D. Melt curve of Wnt5a qPCR on control cells. The melt curve on NMDA-stimulated cells was similar (not shown). E. RT-PCR results of Wnt5a in control and NMDA-treated cells.

Article Snippet: Anti-Wnt5a antibody was purchased from R&D Systems (AF654); anti-p-P70S6K (Thr389) antibody from Cell Signaling Technology (#9206); anti-GAPDH antibody from Santa Cruz (SC-32233); anti-Synapsin I from Millipore (AB1543P); and FITC-conjugated donkey anti-rabbit secondary antibody (711-097-020) and Rhodamine-conjugated rabbit anti-goat secondary antibody (305-297-003) from Jackson.

Techniques: Control, Incubation, Western Blot, Quantitative RT-PCR, Two Tailed Test, Reverse Transcription Polymerase Chain Reaction

Journal: Molecular Brain

Article Title: NMDA receptor activation stimulates transcription-independent rapid wnt5a protein synthesis via the MAPK signaling pathway

doi: 10.1186/1756-6606-5-1

Figure Lengend Snippet: mTOR signaling pathway is not required for the NMDAR-dependent Wnt5a protein synthesis . Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 25nΜ Rapamycin for 30 min, followed by addition of 50 μΜ NMDA for 15 min. Western blotting analysis of Wnt5a and phosphor-P70S6K proteins were performed. Graphs (mean ± SEM) are from four independent experiments (*, p < 0.05; **, p < 0.01; #, p > 0.05; One-way ANOVA).

Article Snippet: Anti-Wnt5a antibody was purchased from R&D Systems (AF654); anti-p-P70S6K (Thr389) antibody from Cell Signaling Technology (#9206); anti-GAPDH antibody from Santa Cruz (SC-32233); anti-Synapsin I from Millipore (AB1543P); and FITC-conjugated donkey anti-rabbit secondary antibody (711-097-020) and Rhodamine-conjugated rabbit anti-goat secondary antibody (305-297-003) from Jackson.

Techniques: Control, Western Blot

Journal: Molecular Brain

Article Title: NMDA receptor activation stimulates transcription-independent rapid wnt5a protein synthesis via the MAPK signaling pathway

doi: 10.1186/1756-6606-5-1

Figure Lengend Snippet: NMDAR activation stimulates Wnt5a protein synthesis via the MAPK signaling pathway . A. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 20 μM PD98059 (PD98) for 30 min, and then stimulated with 50 μΜ NMDA for 15 min. Wnt5a protein was measured by Western blotting and quantified data were presented in graphs (mean ± SEM; n = 3, *p < 0.05; One-way ANOVA). B. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 20 μM U0126 for 30 min, followed by 50 μΜ NMDA for 15 min. Graphs (mean ± SEM) are from three independent experiments (*, p < 0.05; **, p < 0.01; One-way ANOVA).

Article Snippet: Anti-Wnt5a antibody was purchased from R&D Systems (AF654); anti-p-P70S6K (Thr389) antibody from Cell Signaling Technology (#9206); anti-GAPDH antibody from Santa Cruz (SC-32233); anti-Synapsin I from Millipore (AB1543P); and FITC-conjugated donkey anti-rabbit secondary antibody (711-097-020) and Rhodamine-conjugated rabbit anti-goat secondary antibody (305-297-003) from Jackson.

Techniques: Activation Assay, Control, Western Blot

Journal: Molecular Brain

Article Title: NMDA receptor activation stimulates transcription-independent rapid wnt5a protein synthesis via the MAPK signaling pathway

doi: 10.1186/1756-6606-5-1

Figure Lengend Snippet: A working model for NMDAR-regulated Wnt5a protein synthesis . Stimulation of NMDAR activates MAPK signaling to elicit de novo translation from pre-existing Wnt5a mRNAs.

Article Snippet: Anti-Wnt5a antibody was purchased from R&D Systems (AF654); anti-p-P70S6K (Thr389) antibody from Cell Signaling Technology (#9206); anti-GAPDH antibody from Santa Cruz (SC-32233); anti-Synapsin I from Millipore (AB1543P); and FITC-conjugated donkey anti-rabbit secondary antibody (711-097-020) and Rhodamine-conjugated rabbit anti-goat secondary antibody (305-297-003) from Jackson.

Techniques:

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: (A) Immunohistochemical analysis of human specimens of urothelial carcinoma of the bladder for the expression of Wnt5a, ROR2, CTHRC1 and E-cadherin. Left column, tissue sections from a representative case of low grade urothelial carcinoma; the right column, tissue sections from a representative case of high grade urothelial carcinoma. The middle column represents the trend of expression for each protein in all 15 samples. The expression of Wnt5a, ROR2, and CTHRC1 increases in high grade tumors while E-cadherin shows an opposite trend. Bar=50 μm. (B) Statistical analysis was performed to investigate the correlation between tumor histological grade and immunostaining for Wnt5a, Ror2, CTHRC1, and E-cadherin. Statistical significance was tested at an alpha of 0.05. The software PASW Statistics 18 was used for data analysis (Pearson Education, New York City, NY).

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques: Immunohistochemical staining, Expressing, Immunostaining, Software

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: Wnt5a (A) , Ror2 and CTHRC1 (B) , E-cadherin and vimentin (C) , in RT4, J82 and T24 urothelial carcinoma cell lines.

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques:

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: Top row: H&E stain shows morphological differences between each cell line. Middle row: confocal microscopy images show the expression of Wnt5a (green) for each cell line. Bottom row: confocal microscopy image of the merge expression of Ror2 (green) and CTHRC1 (red) for each cell line. Although the co-expression and colocalization of Ror2 and CTHRC1 is present in all cell lines, it is clearest for RT4 and J82 cell lines.

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques: Staining, Confocal Microscopy, Expressing

Journal: Oncotarget

Article Title: Wnt5a / planar cell polarity signaling pathway in urothelial carcinoma, a potential prognostic biomarker

doi: 10.18632/oncotarget.15877

Figure Lengend Snippet: Primer sequences used for real-time RT-PCR analyses

Article Snippet: To one of each of the two sections the following antihuman primary antibodies were applied: mouse monoclonal antibody against Wnt5a (Abcam, Cambridge, MA) applied at 8.33 μg/ml; rabbit polyclonal antibody against Ror2 (Abnova, Taipei City, Taiwan) applied at 1/100 dilution, and mouse monoclonal antibody against CTHRC1 (Abnova, Taipei City, Taiwan) applied at 4.0 μg/ml.

Techniques: Quantitative RT-PCR, Ubiquitin Proteomics

Journal: Journal of clinical gastroenterology

Article Title: Intramucosal Distribution of WNT Signaling Components in Human Esophagus

doi:

Figure Lengend Snippet: Primers for Real-time PCR

Article Snippet: Wnt 1 and Wnt 5a were raised in goats against a peptide mapping near the N-terminus of Wnt1 of human origin and a peptide mapping within an internal region of Wnt 5a of human origin, respectively (Santa Cruz Biotechnology Inc, Santa Cruz, CA).

Techniques:

Journal: Journal of clinical gastroenterology

Article Title: Intramucosal Distribution of WNT Signaling Components in Human Esophagus

doi:

Figure Lengend Snippet: Real-time PCR and IH analysis of Wnt 5a in human esophageal mucosa. A, Real-time PCR data for Wnt 5a in laser-capture microdissected SC layer, the IC layer, the BC layer, and the LP. Each sample was run in triplicate and the means were normalized with GAPDH to obtain the expression ratio. The means of the expression ratios of Wnt 5a/GAPDH ± SEM of 7 samples are displayed in A. The expression of Wnt 5a was significantly different between the mucosal layers (ANOVA, P<0.03). Wnt 5a expression in the IC layer was significantly greater than the BC layer (*P<0.002, n = 7). B and C, IH of cryosections (10 μm) of full thickness human esophagus. Wnt 5a-specific staining (→) was observed in the IC layer and the SC layer (C). Negative IgG-isotype–matched controls on serial sections are also shown (B). Microphotographs are representative of IH performed on 6 esophagi (×100 magnification).

Article Snippet: Wnt 1 and Wnt 5a were raised in goats against a peptide mapping near the N-terminus of Wnt1 of human origin and a peptide mapping within an internal region of Wnt 5a of human origin, respectively (Santa Cruz Biotechnology Inc, Santa Cruz, CA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Staining