Question: You believe that the cytosolic protein p007 is involved in mediating growth hormone-stimulated proliferation in cells. You wish to...

Question:

You believe that the cytosolic protein p007 is involved in mediating growth hormone-stimulated proliferation in cells. You wish to establish a cell model using your cultured fibroblasts. Provide clear explanations to the following,

a) Describe how you would transfect the growth hormone receptor DNA into your cells to create stable cell lines, and include a description of the benefits of using stable cell lines versus transiently transfected cell line. 

b) Describe how you would measure specific receptor binding of GH to your stable cell line.

c) Describe how you would measure growth hormone-induced proliferation in your stable cell line. Make sure to include controls.

d) Describe how you would use RNAi to demonstrate the requirement for p007 in growth-hormone stimulated proliferation. Make sure to include controls.