Wnt-3a Antibody (930769)

Topflash Reporter Activity Induced by Wnt-3a and Neutralization by Human/Mouse Wnt-3a Antibody.

Recombinant Mouse Wnt-3a (Catalog # 1324-WN) induces Topflash reporter activity in the HEK293T human embryonic kidney cell line in a dose-dependent manner (orange line), as measured by the Topflash Reporter Assay. Under these conditions, Topflash reporter activity elicited by Recombinant Mouse Wnt-3a (0.5 µg/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB9025). The ND₅₀ is typically 0.15-0.9 µg/mL.

Detection of Wnt-3a by Western Blot

STAT3 involvement in the Wnt3a-mediated enhancement of IL-4 response.(A) Proportion of differentially expressed genes (using RNA-seq) of bone marrow-derived macrophages (BMDMs) treated with IL-4 (green), Wnt3a (orange), or their combination (purple) compared to untreated cells. (B) Top eight GO terms of differentially expressed genes in Wnt3a and Wnt3a+IL-4-treated BMDMs. (C) Representative western blot image for phosphorylates (pY) and total STAT3 following treatment of BMDMs with Wnt3a for the indicated times (one out of two independent experiments). (D) BMDMs used in Figure 7F were examined for STAT3 expression using qPCR. (E) Representative western blot image for phosphorylates and total STAT3, as well as GAPDH, following treatment of BMDMs with Wnt3a or its conditioned media (CM) for the indicated times (one out of three independent experiments). Wnt CM was produced by treatment of BMDMs with Wnt3a for 16 hr. (F) BMDMs were treated with Wnt3a for 16 hr and added IL-4 for 2.5 hr thereafter, or with CM+IL-4 for 2.5 hr. Some CM was treated for 1 hr with a blocking antibody to Wnt3a prior to their exposure (in combination with IL4) to naïve BMDMs for 2.5 hr. Arg1 expression was determined with qPCR. p-Values were determined via one-way ANOVA, compared with IL-4 treatment, and Dunnett’s multiple comparison test (with repeated measurements). (G) BMDMs were treated with IL-4, with or without Wnt3a, and with blocking antibodies for transforming growth factor beta, C-X-C motif chemokine receptor 3, glycoprotein 103, IL-10 receptor, and hepatocyte growth factor receptor for 16 hr, and Arg1 expression was determined using real-time qPCR (n = 2–3 mice per treatment). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33720008), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Wnt-3a by Western Blot

STAT3 involvement in the Wnt3a-mediated enhancement of IL-4 response.(A) Proportion of differentially expressed genes (using RNA-seq) of bone marrow-derived macrophages (BMDMs) treated with IL-4 (green), Wnt3a (orange), or their combination (purple) compared to untreated cells. (B) Top eight GO terms of differentially expressed genes in Wnt3a and Wnt3a+IL-4-treated BMDMs. (C) Representative western blot image for phosphorylates (pY) and total STAT3 following treatment of BMDMs with Wnt3a for the indicated times (one out of two independent experiments). (D) BMDMs used in Figure 7F were examined for STAT3 expression using qPCR. (E) Representative western blot image for phosphorylates and total STAT3, as well as GAPDH, following treatment of BMDMs with Wnt3a or its conditioned media (CM) for the indicated times (one out of three independent experiments). Wnt CM was produced by treatment of BMDMs with Wnt3a for 16 hr. (F) BMDMs were treated with Wnt3a for 16 hr and added IL-4 for 2.5 hr thereafter, or with CM+IL-4 for 2.5 hr. Some CM was treated for 1 hr with a blocking antibody to Wnt3a prior to their exposure (in combination with IL4) to naïve BMDMs for 2.5 hr. Arg1 expression was determined with qPCR. p-Values were determined via one-way ANOVA, compared with IL-4 treatment, and Dunnett’s multiple comparison test (with repeated measurements). (G) BMDMs were treated with IL-4, with or without Wnt3a, and with blocking antibodies for transforming growth factor beta, C-X-C motif chemokine receptor 3, glycoprotein 103, IL-10 receptor, and hepatocyte growth factor receptor for 16 hr, and Arg1 expression was determined using real-time qPCR (n = 2–3 mice per treatment). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33720008), licensed under a CC-BY license. Not internally tested by R&D Systems.