Human Frizzled-6 Antibody

Frizzled‑6 in Human Kidney Cancer Tissue. Frizzled-6 was detected in immersion fixed paraffin-embedded sections of human kidney cancer tissue using Goat Anti-Human Frizzled-6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3149) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; (CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Frizzled-6 by Flow Cytometry miR-20b and miR-125b augment Wnt signalling through targeting FZD6. (h) sphere formation of MES 83 expressing control shRNA or shRNAs of FZD6, with or without re-expression of FZD6, or a control (right). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27698350), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Frizzled-6 by Western Blot miR-20b and miR-125b augment Wnt signalling through targeting FZD6. (d) IB for FZD6 and APC in miR-transduced MES 83 and 1123, or miRZIP-transduced PN 157 and 84. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27698350), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Frizzled-6 by Western Blot FZD6 mediates induction of the MES-associated phenotype and suppression of the Wnt signalling through TAK1.(a) IB analysis on PN 23 and 17 spheres with or without FZD6 overexpression using indicated antibodies. (b) qRT–PCR analysis of PN-associated genes OLIG2 and SOX2, and MES-associated genes ALDH1A3 and CD44 in PN 23 spheres with or without FZD6 overexpression. (c) FZD6 expression promoted PN 23 spheres proliferation and self-renewal. (d) Ectopic expression of FZD6 promoted tumour growth of PN 23 spheres in GBM xenografts in the brain (n=5). Scale bar, 1.0 mm. (e) TOPFlash/FOPFlash assays on PN 23 with overexpressing FZD6 or control, with or without CaMKII inhibitor KN93 (5 μM), TAK1 inhibitor 5Z-7-oxo (5 μM), or a shRNA-NLK. (f,g) qRT–PCR analyses of the relative expression of miR-20b and miR-125b (f), and Wnt signalling target gene LEF1 (g) in PN 23 overexpressing FZD6, or a vector control, with or without CaMKII inhibitor KN93 (5 μM), TAK1 inhibitor 5Z-7-oxo (5 μM), or a shRNA-NLK. (h) sphere formation assays for PN 23 and 17 spheres with or without overexpression of FZD1, FZD3, and FZD5, respectively. (i) Summary of data shown in Supplementary Fig. 10. An overview of transmembrane receptors and Wnt ligands that affect FZD6 overexpression-induced phenotypes. Data were normalized to the control. *Relative inhibitory effects on indicated protein expression or cellular properties. Error bars (s.d.) represent the data of triplicate samples for each cell line or tumours from five individual mice in each group. *P<0.05, **P<0.01, paired two-way Student's t-test. Data are representative from three independent experiments with similar results. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27698350), licensed under a CC-BY license. Not internally tested by R&D Systems.