Quantitative Live-cell Reporter Assay for Noncanonical Wnt Activity.

Bio-protocol
Edith P KarunaHsin-Yi Henry Ho

Abstract

Noncanonical Wnt signaling functions independently of the β-catenin pathway to control diverse developmental processes, and dysfunction of the pathway contributes to a number of human pathological conditions, including birth defects and metastatic cancer. Progress in the field, however, has been hampered by the scarcity of functional assays for measuring noncanonical Wnt signaling activity. We recently described the Wnt5a-Ror-Kif26b (WRK) reporter assay, which directly monitors a post-transcriptional regulatory event in noncanonical Wnt signaling. In this protocol, we describe the generation of the stable GFP-Kif26b reporter cell line and a quantitative reporter assay for detecting and measuring Wnt5a signaling activities in live cells via flow cytometry.

References

Feb 27, 2010·PloS One·Christophe Fuerer, Roel Nusse
May 12, 2010·Molecular and Cellular Biology·Michiru NishitaYasuhiro Minami
Dec 4, 2010·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Bisei Ohkawara, Christof Niehrs
Feb 22, 2012·Proceedings of the National Academy of Sciences of the United States of America·Hsin-Yi Henry HoMichael Eldon Greenberg

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Citations

Jun 18, 2021·Proceedings of the National Academy of Sciences of the United States of America·Sara E Konopelski SnavelyHsin-Yi Henry Ho

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Methods Mentioned

BETA
flow cytometry
Transfection

Software Mentioned

GenJet
FlowJo

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