what cell line to use - to demonstrate ubiquitination (Apr/12/2011 )

Hi,

My gene of interest shows tumor suppressor properties in vitro and also reduced tumorigenicity in vivo. I did these experiemnts using stable cell lines.

Now I want to check if my gene of interest reduces beta catenin via ubiquitination. I'm planning to treat my cells with a proteosome inhibitor and then check for beta catenin expression in the presence of my gene of interest. But I'm not sure which cell line to use? Do I demonstrate this in my stable cancer cell line which overexpresses my gene of interest or do I demonstrate this in the same cancer cell line but transiently or do i demonstrate this in HK293 cells?

Any advice would be appreciated.

Any of the cells should be fine - though I would use more than one cell type to confirm if you are planning on using your stable cell line.