I'm looking at setting up scratch wound migration experiments, this normally takes 24-48hs. I am using MCF10A stable cell lines and MDA-MB-231 cells with transient transfection against the same set of genes. Scratch wound assays I've performed have given me a difference relative to my negative controls, and now I want to test whether this is sensitive to EGF induction.
In my reading, I've only seen EGF induction for a maximum of 1hr, I believe if it is prolonged then negative feedback gets in the way. My worry is that if I only treat for 1 hour then the effects won't carry through for the full time course.
My question is, how can I have a prolonged EGF induction over my 24-48 hour time course?
An additional minor question, after serum starvation, do you add EGF and serum or just EGF?
Looking forward to some enlightenment