rabbit anti fzd4 antibody  (Bioss)

Journal: Frontiers in Physiology

Article Title: In Ovo Injection of CHIR-99021 Promotes Feather Follicle Development via Modulating the Wnt Signaling Pathway and Transcriptome in Goose Embryos ( Anser cygnoides )

doi: 10.3389/fphys.2022.858274

Figure Lengend Snippet: In ovo injection of CHIR-99021 in goose embryos activates the Wnt signaling pathway by inhibiting GSK-3β. The expression of FZD4, β-catenin, LEF1, TCF4, and c-Myc in dorsal tissues of each group at E13 (A) , E17 (B) , E21 (C), and E25 (D) , as detected by western blotting. All data are shown as mean ± SEM, no common superscript letter within the same embryonic day means statistical difference ( p < 0.05) by one-way ANOVA with Duncan’s multiple-range test. n = 6 embryos.

Article Snippet: The antibodies identifiers and dilutions were as follows: rabbit anti-β-catenin antibody (Cat# bs-23663R, bioss, Beijing, China), rabbit anti-GSK-3 β antibody (Cat# bs-0023M, bioss), 1:2,000,1:2,000; rabbit anti-c-Myc antibody (Cat# bs-4963R, bioss), 1:2,000; rabbit anti-TCF4 antibody (Cat# bs-1280R, bioss), 1:2000; rabbit anti-LEF1 antibody (Cat# bs-1843R, bioss), 1:1,500; rabbit anti-FZD4 antibody (Cat# bs-13217R, bioss), 1:2,000; ACTB rabbit monoclonal antibody (Cat# AC026, Abclonal, Wuhan, China), 1:50,000.

Techniques: In Ovo, Injection, Expressing, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: GATA6-AS1 Is Poorly Expressed and FZD4 Is Highly Expressed in GC (A) Heatmap of GEO: GSE13911 . (B) Heatmap of GEO: GSE19826 . (C) FZD4 expression in GC from TCGA, with red boxplot referring to GC sample and gray boxplot referring to normal sample. (D) Survival analysis of FZD4 in GC from TCGA, with red boxplot referring to patients with high expression of FZD4 and gray boxplot referring to patients with low expression of FZD4. (E) Methylation of FZD4 in different stages of GC. (F) Expression of FZD4 in GC tissues and paracancerous normal tissues examined by immunohistochemistry (original magnification, ×400). *p < 0.05 versus the paracancerous normal tissues. (G) mRNA and protein levels of GATA6-AS1 and FZD4 in CG tissues determined by qRT-PCR and western blot analysis. *p < 0.05 versus the paracancerous normal tissues. (H) Correlation analysis of GATA6-AS1 and FZD4. (I) Expression of GATA6-AS1 and mRNA and protein level of FZD4 in gastric cell line and GC cell lines. *p < 0.05 versus GES-1 cell line. In (A) and (B), the abscissa represents the sample number, the ordinate indicates the name of the differentially expressed genes, and the upper right histogram is the color gradation. The color change from top to bottom indicates the decrease of expression value, and each rectangle corresponds to the expression value of a sample. Each column represents the expression of all genes in each sample. The left dendrogram represents the results of cluster analysis of different genes from different samples. The top cross bar shows the sample type, and the upper right box reveals the sample color reference. The blue is the normal control sample, and the red is the tumor sample.

Article Snippet: After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c- myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3β Ser9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA).

Techniques: Expressing, Methylation, Immunohistochemistry, Quantitative RT-PCR, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: KEGG Pathway Enrichment Analysis of the Target Gene of GATA6-AS1

Article Snippet: After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c- myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3β Ser9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA).

Techniques: Protein-Protein interactions

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: Association between GATA6-AS1, FZD4 Expression, and Clinicopathological Features

Article Snippet: After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c- myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3β Ser9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA).

Techniques: Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: FZD4 Binds Specifically to GATA6-AS1 (A) Distribution of GATA6-AS1 expression in cells predicted by bioinformatic website lncATLAS. (B) Distribution of GATA6-AS1 expression in cells detected by FISH assay (original magnification, 400). (C) Binding site between GATA6-AS1 and FZD4 by bioinformatics prediction. (D) Relative luciferase activity measured by dual-luciferase reporter gene assay. *p < 0.05 versus the NC group. (E) mRNA expression of GATA6-AS1 and FZD4 determined by qRT-PCR. *p < 0.05 versus the blank group. (F and G) Protein levels of GATA6-AS1 and FZD4 normalized to GAPDH determined by western blot analysis. *p < 0.05 versus the blank group. (H) Relative GATA6-AS1 levels detected by RIP assay. *p < 0.05 versus the IgG group. (I) Protein level of FZD4 normalized to GAPDH determined by western blot analysis. *p < 0.05 versus the si-NC group. (J) Enrichment of H3K27me3 and EZH2 relative to input in the FZD4 promoter region detected by ChIP assay. *p < 0.05 versus the NC group.

Article Snippet: After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c- myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3β Ser9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA).

Techniques: Expressing, Binding Assay, Luciferase, Activity Assay, Reporter Gene Assay, Quantitative RT-PCR, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: The Molecular Mechanism Involving the Role of GATA6-AS1 and FZD4 in GC Development via the Wnt/β-Catenin Signaling Pathway Overexpression of GATA6-AS1 downregulated the expression of FZD4 by recruiting EZH2 and H3K27me3 in the FZD4 promoter region to inactivate the Wnt/β-catenin signaling pathway, thus inhibiting proliferation, EMT, and lymphatic metastasis in GC.

Article Snippet: After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c- myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3β Ser9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA).

Techniques: Over Expression, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: Primer Sequences for qRT-PCR

Article Snippet: After the membrane had been blocked for 1 h, it was incubated with primary antibodies of rabbit polyclonal antibodies to FZD4 (1 μg/mL, ab83042), β-catenin (1:4,000, ab6302), c- myc (1:500, ab39688), cyclinD1 (1:500, ab61758), E-cadherin (1:500, ab15148), Vimentin (1:1,000, ab137321), Snail (1:500, ab82846), Slug (1:100, ab75629), p-GSK-3β Ser9 (1:500, ab131097), and mouse monoclonal antibody to GSK-3β (1:500, ab93926) overnight at 4°C (all antibodies above were purchased from Abcam, Cambridge, MA, USA).

Techniques: Sequencing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: GATA6-AS1 Is Poorly Expressed and FZD4 Is Highly Expressed in GC (A) Heatmap of GEO: GSE13911 . (B) Heatmap of GEO: GSE19826 . (C) FZD4 expression in GC from TCGA, with red boxplot referring to GC sample and gray boxplot referring to normal sample. (D) Survival analysis of FZD4 in GC from TCGA, with red boxplot referring to patients with high expression of FZD4 and gray boxplot referring to patients with low expression of FZD4. (E) Methylation of FZD4 in different stages of GC. (F) Expression of FZD4 in GC tissues and paracancerous normal tissues examined by immunohistochemistry (original magnification, ×400). *p < 0.05 versus the paracancerous normal tissues. (G) mRNA and protein levels of GATA6-AS1 and FZD4 in CG tissues determined by qRT-PCR and western blot analysis. *p < 0.05 versus the paracancerous normal tissues. (H) Correlation analysis of GATA6-AS1 and FZD4. (I) Expression of GATA6-AS1 and mRNA and protein level of FZD4 in gastric cell line and GC cell lines. *p < 0.05 versus GES-1 cell line. In (A) and (B), the abscissa represents the sample number, the ordinate indicates the name of the differentially expressed genes, and the upper right histogram is the color gradation. The color change from top to bottom indicates the decrease of expression value, and each rectangle corresponds to the expression value of a sample. Each column represents the expression of all genes in each sample. The left dendrogram represents the results of cluster analysis of different genes from different samples. The top cross bar shows the sample type, and the upper right box reveals the sample color reference. The blue is the normal control sample, and the red is the tumor sample.

Article Snippet: The sections were subsequently blocked with 3% BSA for 20–30 min at 37°C, incubated with primary antibody of rabbit polyclonal antibody to FZD4 (1:500, ab80049, Abcam, Cambridge, MA, USA) for 2 h at 37°C, and further incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000, Zhongshan Bio-Tech, Guangzhou, Guangdong, China) in a wet box for 30 min at 37°C.

Techniques: Expressing, Methylation, Immunohistochemistry, Quantitative RT-PCR, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: KEGG Pathway Enrichment Analysis of the Target Gene of GATA6-AS1

Article Snippet: The sections were subsequently blocked with 3% BSA for 20–30 min at 37°C, incubated with primary antibody of rabbit polyclonal antibody to FZD4 (1:500, ab80049, Abcam, Cambridge, MA, USA) for 2 h at 37°C, and further incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000, Zhongshan Bio-Tech, Guangzhou, Guangdong, China) in a wet box for 30 min at 37°C.

Techniques: Protein-Protein interactions

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: Association between GATA6-AS1, FZD4 Expression, and Clinicopathological Features

Article Snippet: The sections were subsequently blocked with 3% BSA for 20–30 min at 37°C, incubated with primary antibody of rabbit polyclonal antibody to FZD4 (1:500, ab80049, Abcam, Cambridge, MA, USA) for 2 h at 37°C, and further incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000, Zhongshan Bio-Tech, Guangzhou, Guangdong, China) in a wet box for 30 min at 37°C.

Techniques: Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: FZD4 Binds Specifically to GATA6-AS1 (A) Distribution of GATA6-AS1 expression in cells predicted by bioinformatic website lncATLAS. (B) Distribution of GATA6-AS1 expression in cells detected by FISH assay (original magnification, 400). (C) Binding site between GATA6-AS1 and FZD4 by bioinformatics prediction. (D) Relative luciferase activity measured by dual-luciferase reporter gene assay. *p < 0.05 versus the NC group. (E) mRNA expression of GATA6-AS1 and FZD4 determined by qRT-PCR. *p < 0.05 versus the blank group. (F and G) Protein levels of GATA6-AS1 and FZD4 normalized to GAPDH determined by western blot analysis. *p < 0.05 versus the blank group. (H) Relative GATA6-AS1 levels detected by RIP assay. *p < 0.05 versus the IgG group. (I) Protein level of FZD4 normalized to GAPDH determined by western blot analysis. *p < 0.05 versus the si-NC group. (J) Enrichment of H3K27me3 and EZH2 relative to input in the FZD4 promoter region detected by ChIP assay. *p < 0.05 versus the NC group.

Article Snippet: The sections were subsequently blocked with 3% BSA for 20–30 min at 37°C, incubated with primary antibody of rabbit polyclonal antibody to FZD4 (1:500, ab80049, Abcam, Cambridge, MA, USA) for 2 h at 37°C, and further incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000, Zhongshan Bio-Tech, Guangzhou, Guangdong, China) in a wet box for 30 min at 37°C.

Techniques: Expressing, Binding Assay, Luciferase, Activity Assay, Reporter Gene Assay, Quantitative RT-PCR, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: The Molecular Mechanism Involving the Role of GATA6-AS1 and FZD4 in GC Development via the Wnt/β-Catenin Signaling Pathway Overexpression of GATA6-AS1 downregulated the expression of FZD4 by recruiting EZH2 and H3K27me3 in the FZD4 promoter region to inactivate the Wnt/β-catenin signaling pathway, thus inhibiting proliferation, EMT, and lymphatic metastasis in GC.

Article Snippet: The sections were subsequently blocked with 3% BSA for 20–30 min at 37°C, incubated with primary antibody of rabbit polyclonal antibody to FZD4 (1:500, ab80049, Abcam, Cambridge, MA, USA) for 2 h at 37°C, and further incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000, Zhongshan Bio-Tech, Guangzhou, Guangdong, China) in a wet box for 30 min at 37°C.

Techniques: Over Expression, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Overexpressed lncRNA GATA6-AS1 Inhibits LNM and EMT via FZD4 through the Wnt/β-Catenin Signaling Pathway in GC

doi: 10.1016/j.omtn.2019.09.034

Figure Lengend Snippet: Primer Sequences for qRT-PCR

Article Snippet: The sections were subsequently blocked with 3% BSA for 20–30 min at 37°C, incubated with primary antibody of rabbit polyclonal antibody to FZD4 (1:500, ab80049, Abcam, Cambridge, MA, USA) for 2 h at 37°C, and further incubated with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:1,000, Zhongshan Bio-Tech, Guangzhou, Guangdong, China) in a wet box for 30 min at 37°C.

Techniques: Sequencing