If you understand well the activation of Wnt signaling, the signaling requires the inhibition of beta-catenin destruction complex and the localization of beta-catenin to the nucleus before interacting with adaptor proteins and transcription factors to transcribe Wnt target genes. Based on this information, you can check whether Wnt signaling is active by checking whether beta-catenin has successfully translocated into the nucleus. This can easily be observed by immunofluorescence assay that stains beta-catenin protein and DNA stain to mark the nuclear compartment. Additionally, you can check on the expression of Wnt target genes by using RT-qPCR or western blotting. By looking at both the translocation of beta-catenin and the expression of Wnt target genes, you should be able to get strong confirmation of whether the pathway is blocked or activated.

When assessing Wnt activator and Wnt inhibitor, be sure to have different cell line that has sufficiently low Wnt activity (when assessing activator) or sufficiently high Wnt activity (when assessing inhibitor). In my studies, I studied compounds that inhibit Wnt activity by artificially activating the Wnt pathway using the GSK3 inhibitor.

I hope this helps. Good luck!

Thank you so much Sir. Win Sen Heng

Will you help me to how can I perform this immunofluorescence assay ? Any reference article which will help me.

Can you share me your article so, I can download it and refer it.

Immunofluorescence assay can be performed by using an antibody to bind to your target protein and using a secondary antibody-conjugated with a flurophore to resolve the signal. A simple online search can give you a good protocol. As for my own paper, you can check my profile. There are two papers related to beta-catenin; both of which use immunofluorescence assay to detect localization of beta-catenin.

Thank you so much sir.

I view your profile and get the article.

Thank you so much once again.

It will be really helpful for me in my study. Win Sen Heng