primary antibodies against wnt3a  (Santa Cruz Biotechnology)

Journal: Stem Cell Research & Therapy

Article Title: A small molecule modulating monounsaturated fatty acids and Wnt signaling confers maintenance to induced pluripotent stem cells against endodermal differentiation

doi: 10.1186/s13287-021-02617-x

Figure Lengend Snippet: Analysis of the Wnt signaling pathway following endoderm differentiation of pluripotent stem cells. The induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) were treated with SCD1 inhibitor (SCDinhib) alone at Days 0, 1, or 2 (0, 1, and 2) of differentiation or in combination with oleic acid (OA) at Day 0. A The representative Western blot and the quantification of Wnt3a and β-catenin. * p < 0.05 versus mock, # p < 0.05, ## p < 0.01 versus the same Day in SCDinhib

Article Snippet: Primary antibodies against Wnt3a (Santa Cruz, USA) and β-catenin (Santa Cruz, USA) were applied for evaluation of the Wnt signaling pathway.

Techniques: Western Blot

Journal: BMC Gastroenterology

Article Title: In-depth characterization of the Wnt-signaling/β-catenin pathway in an in vitro model of Barrett’s sequence

doi: 10.1186/s12876-019-0957-5

Figure Lengend Snippet: Oligo sequences

Article Snippet: Diluted 1:50 in 1% BSA in PBS, the monoclonal primary antibody against WNT3A (AM09053PU-N, OriGene Technologies GmbH, Herford, Germany) was incubated over night at 4 °C.

Techniques:

Journal: BMC Gastroenterology

Article Title: In-depth characterization of the Wnt-signaling/β-catenin pathway in an in vitro model of Barrett’s sequence

doi: 10.1186/s12876-019-0957-5

Figure Lengend Snippet: Expression of the extracellular ligands Wnt3a and Wnt5a along Barrett’s sequence. Expression of Wnt3a ( a ) and Wnt5a ( b ) was analyzed in EPC-1 and EPC-2 (squamous esophageal epithelium), CP-A (metaplastic epithelium), CP-B (epithelium with high-grade dysplasia), OE33 and OE19 (esophageal adenocarcinoma) cells by quantitative Real time RT-PCR. Highest Wnt3a expression was detected in EPC-1 and EPC-2 ( a ). Wnt5a was also expressed in EPC-1 and EPC-2, but highest levels of Wnt5a were found in CP-B ( b ). OE33 and OE19 expressed only marginal levels of Wnt3a and Wnt5a ( a , b ). Normalization was done with β-Actin. Values are shown as mean ± S.E.M. (One-way-ANOVA with Bonferroni correction, * - p < 0.05, ** - p < 0.01, *** - p < 0.001 compared to EPC-1)

Article Snippet: Diluted 1:50 in 1% BSA in PBS, the monoclonal primary antibody against WNT3A (AM09053PU-N, OriGene Technologies GmbH, Herford, Germany) was incubated over night at 4 °C.

Techniques: Expressing, Sequencing, Quantitative RT-PCR

Journal: BMC Gastroenterology

Article Title: In-depth characterization of the Wnt-signaling/β-catenin pathway in an in vitro model of Barrett’s sequence

doi: 10.1186/s12876-019-0957-5

Figure Lengend Snippet: Expression pattern of different Wnt-signaling molecules in patients’ biopsies from squamous epithelium, Barrett’s mucosa, HGIN and EAC. Expression of the receptors Fzd1–10, the co-receptors LRP5/6 and ligands Wnt3a and Wnt5a were analyzed by RT-PCR in biopsies taken during endoscopy from squamous epithelium (SQ_1–7) and their corresponding Barrett’s mucosa (BM_1–7), as well as in specimens from patients with high-grade intraepithelial neoplasia (HGIN_1–7) and esophageal adenocarcinoma (EAC_1–7). Biopsies were placed directly in RNAlater®. PCR products were applied to 2% ethidiumbromid stained agarose and visualized with UV light

Article Snippet: Diluted 1:50 in 1% BSA in PBS, the monoclonal primary antibody against WNT3A (AM09053PU-N, OriGene Technologies GmbH, Herford, Germany) was incubated over night at 4 °C.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Staining

Journal: BMC Gastroenterology

Article Title: In-depth characterization of the Wnt-signaling/β-catenin pathway in an in vitro model of Barrett’s sequence

doi: 10.1186/s12876-019-0957-5

Figure Lengend Snippet: Response to Wnt3a in cells of Barrett’s sequence. Expression of the downstream target Axin2 was analyzed in EPC-1 and EPC-2, CP-A, CP-B, OE33 and OE19 cells by quantitative Real time RT-PCR ( a ). The carcinoma cell lines OE33 and OE19 were treated with 200 ng/mL rhWnt3a for 1 h. Cell specific FCS free medium served as control. Expression of the downstream targets Axin2 and CyclinD1 were analyzed by quantitative Real time RT-PCR ( b , c ). Along Barrett’s sequence, a strong increase of Axin2 expression was found with significant higher levels in OE19 as compared to EPC-1 ( a ). EPC-1, CP-A and OE33 cells responded to Wnt3a with an increased Axin2 expression. In contrast, EPC-2 and OE19 showed no altered Axin2 expression as compared to controls ( b ). Decreased levels of CyclinD1 after Wnt3a were detected in EPC-1, EPC-2, CP-A, and CP-B. Wnt3a treatment induced significant higher CyclinD1 levels in OE33 cells as compared to controls ( c ). Protein level expression of pAkt/Akt, pβ-catenin/β-catenin and pGSK3β/GSK3β after Wnt3a treatment was analyzed in EPC-1 and EPC-2, CP-A, CP-B, OE33 and OE19 cells by westernblot analyses. Higher levels of pβ-catenin after Wnt3a treatment were found in OE33 cells ( d ). CP-B responded with an elevated pGSK3β expression ( e ). A loss of pAkt was detected in CP-A and CP-B ( f ). Normalization was done with β-Actin. Values are shown as mean ± S.E.M. (One-way-ANOVA with Bonferroni correction, **- p < 0.01 compared to EPC-1 ( a ), Two-way-ANOVA with Bonferroni correction, * - p < 0.05, ** - p < 0.01 compared to controls ( b - f ))

Article Snippet: Diluted 1:50 in 1% BSA in PBS, the monoclonal primary antibody against WNT3A (AM09053PU-N, OriGene Technologies GmbH, Herford, Germany) was incubated over night at 4 °C.

Techniques: Sequencing, Expressing, Quantitative RT-PCR, Control

Journal: BMC Gastroenterology

Article Title: In-depth characterization of the Wnt-signaling/β-catenin pathway in an in vitro model of Barrett’s sequence

doi: 10.1186/s12876-019-0957-5

Figure Lengend Snippet: Immunohistochemically staining of WNT3A in metaplasia, HGIN and EAC. Immunohistochemical staining of WNT3A was performed and the staining index of WNT3A-positive cells ( a ) of squamous epithelial (SQ), metaplastic (MC), high-grade intraepithelial neoplasia (HGIN), and esophageal adenocarcinoma (EAC) cells in human specimens ( b , c ) were analyzed. The staining index used, describes staining as 1 < 30%, 2 < 60%, 3 < 100%. WNT3A protein localization, showed a higher staining index for SQ than MC, HGIN and EAC. Values are shown as mean ± S.E.M. (Kruskal-Wallis with Dunn’s correction, * - p < 0.05, *** - p < 0.001, n SQ = 30, n MC = 15, n HGIN = 7, n EAC = 15, bar-20 μm)

Article Snippet: Diluted 1:50 in 1% BSA in PBS, the monoclonal primary antibody against WNT3A (AM09053PU-N, OriGene Technologies GmbH, Herford, Germany) was incubated over night at 4 °C.

Techniques: Staining, Immunohistochemical staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Combined effects of fibroblast growth factor 2 and dexamethasone on differentiation of human cementoblasts

doi:

Figure Lengend Snippet: Oligonucleotide primer sequences utilized in RT-PCR

Article Snippet: The membranes were blocked for 1 hour with 5% non-fat dry milk in Tris-buffered saline/Tween-20 (TBST) buffer and incubated with primary antibodies against Wnt3a, Runx2, β-catenin, and p-GSK-3β (1:1000 dilution) (Cell Signaling Technology, Inc., Danvers, MA) at 4°C overnight.

Techniques:

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Combined effects of fibroblast growth factor 2 and dexamethasone on differentiation of human cementoblasts

doi:

Figure Lengend Snippet: Combined effect of FGF-2 and dexamethasone on calcium deposition and mRNA expression of cementoblastic markers. Alizarin red S staining of calcium deposition at 21 d in CON group (A) and F2+D2 group (B). *P<0.05. RT-PCR analysis of RUNX2 (C), OCN (D), CAP (E) and BSP (F) mRNA expression at 14 d. *P<0.05.

Article Snippet: The membranes were blocked for 1 hour with 5% non-fat dry milk in Tris-buffered saline/Tween-20 (TBST) buffer and incubated with primary antibodies against Wnt3a, Runx2, β-catenin, and p-GSK-3β (1:1000 dilution) (Cell Signaling Technology, Inc., Danvers, MA) at 4°C overnight.

Techniques: Expressing, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Combined effects of fibroblast growth factor 2 and dexamethasone on differentiation of human cementoblasts

doi:

Figure Lengend Snippet: Combined effects of FGF-2 and dexamethasone on Wnt pathway in cementoblasts and the effects of DKK1. A: Protein expression of Wnt3a, Runx2, β-catenin, and p-GSK-3β were determined by Western blot. B: Western blot analysis of the DKK1 effect on the expression of Wnt pathway molecules. *P<0.05.

Article Snippet: The membranes were blocked for 1 hour with 5% non-fat dry milk in Tris-buffered saline/Tween-20 (TBST) buffer and incubated with primary antibodies against Wnt3a, Runx2, β-catenin, and p-GSK-3β (1:1000 dilution) (Cell Signaling Technology, Inc., Danvers, MA) at 4°C overnight.

Techniques: Expressing, Western Blot