Rhwnt3a

Manufactured by Thermo Fisher Scientific

RhWNT3A is a recombinant human Wnt3a protein. It is used in cell biology research applications.

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Lab products found in correlation

Chlorpromazine hcl, by Merck Group (1 mentions) Wnt3a, by American Type Culture Collection (1 mentions) Hek293t 17, by American Type Culture Collection (1 mentions) Neomycin trisulfate salt hydrate, by Merck Group (1 mentions) Rhwnt5a, by R&D Systems (1 mentions) Chir99021, by Cayman Chemical (1 mentions) Bafilomycin a1, by Cayman Chemical (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 phalloidin, by Thermo Fisher Scientific (1 mentions) Goat anti mouse dylight 488, by Abcam (1 mentions)

2 protocols using rhwnt3a

Cell Culture and Treatment Protocols

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All cells were cultured at 37°C and 5% CO₂. HT1080 (human male), HEK293T/17 (human fetus), RKO (human, gender N/A), Lcells (mouse male), and WNT3A expressing Lcells (mouse male) were obtained from American Type Culture Collection (ATCC, Manassas, VA) and grown in DMEM with 10% FBS. Each cell line was tested for mycoplasma contamination and passaged no more than 20 passages from the original ATCC stock. Cells were treated at the indicated concentrations with the following compounds: CHIR99021 (Cayman Chemicals, Ann Arbor, MI), chlorpromazine HCL (Sigma-Aldrich, St. Louis, MO), bafilomycin A1 (Cayman Chemicals), rhWNT3A (PeproTech, Rocky Hill, NJ), rhWNT5A (R&D, Minneapolis, MN), neomycin trisulfate salt hydrate (Sigma-Aldrich) and carbachol (EMD Chemicals, Gibbstown, NJ).

Agajanian M.J., Walker M.P., Axtman A.D., Ruela-de-Sousa R.R., Serafin D.S., Rabinowitz A.D., Graham D.M., Ryan M.B., Tamir T., Nakamichi Y., Gammons M.V., Bennett J.M., Couñago R.M., Drewry D.H., Elkins J.M., Gileadi C., Gileadi O., Godoi P.H., Kapadia N., Müller S., Santiago A.S., Sorrell F.J., Wells C.I., Fedorov O., Willson T.M., Zuercher W.J, & Major M.B. (2019). WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop. Cell Reports, 26(1), 79-93.e8.

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Quantifying Focal Adhesion Dynamics

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Here, 50,000 cells were seeded on 1 cm Ø glass cover slips in 12-well plates and treated with formate (1 or 10 mM) or rhWnt3A (100 ng ml⁻¹, PeproTech) for 48 h. Cells were fixed in 3.5%PFA for 30 min, washed with PBS and PBS-Trition 1%, blocked with 5% milk and washed again in PBS-Triton 0.5%. Focal adhesions were stained overnight with mouse-anti-Vinculin (primary antibody; V4505, Sigma-Aldrich), before being further washed with PBS-Triton 0.5% and stained with goat-anti-mouse-DyLight488 (ab96879, Abcam), Phalloidin AlexaFluor594 (A12381, ThermoFisher) and DAPI (ThermoFisher). Pictures were randomly taken under a confocal microscope and images were analysed using ImageJ (version 1.53K).

Ternes D., Tsenkova M., Pozdeev V.I., Meyers M., Koncina E., Atatri S., Schmitz M., Karta J., Schmoetten M., Heinken A., Rodriguez F., Delbrouck C., Gaigneaux A., Ginolhac A., Nguyen T.T., Grandmougin L., Frachet-Bour A., Martin-Gallausiaux C., Pacheco M., Neuberger-Castillo L., Miranda P., Zuegel N., Ferrand J.Y., Gantenbein M., Sauter T., Slade D.J., Thiele I., Meiser J., Haan S., Wilmes P, & Letellier E. (2022). The gut microbial metabolite formate exacerbates colorectal cancer progression. Nature Metabolism, 4(4), 458-475.

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