GFP analysis and lab help - (Dec/05/2012 )

How would you would quantitate the average number of cells with nuclear localised GFP signal?
How could you quantitate the average proportion of total GFP that is localised to the nucleus per cell

How could a cell maintain a cytoplasmically localised transcription factor such that after a stimulus this factor would relocate to the nucleus?

How could you use recombinant Ran protein to purify multiple nuclear import factors and iclude two control incubations to ensure the factors isolated are specific Ran binding partners?

You shouldn't get any answers here without providing some input yourself, this is to help you learn rather than us just telling you stuff for you to copy and paste...

Hint: transporters.

I have got answers but just wanted clarification of ideas for example I believe the 3rd question relates to transcription factors like the Nf-kB family which remain in the cytoplasm until activated by phosphorylation of lKB which unmasks the NLS s of p50 and p65 resulting in their translocation into the nucleus.