You have isolated an epithelial cell line which is unresponsive to EGF (normal epithelial derived cell lines require EGF in the culture media for growth). You suspect that EGF receptor signaling in this cell line is impaired, and you designate this a mutant cell line. As an assay for EGFR signaling, you have stably transfected this mutant cell line and a normal cell line with a luciferase-based reporter construct that is responsive to the transcription factor Elk-1. You want to determine what component of the signaling pathway is impaired.
Briefly describe a method you could use to transfect the luciferase-based reporter construct DNA into a cell, and then describe how you could establish (or select for) a stable cell line expressing this reporter construct.
You have isolated an epithelial cell line which is unresponsive to EGF (normal epithelial derived cell...
5. (12pts) You have discovered a novel cell-signaling pathway, in which cells stimulated with a hormone, Simpson, leads to a cell death response. The pathway involves 5 proteins with the following signaling: Simpson (signal) Homer --I Marge > Bart Lisa MaggieCell growth (response) A. Do you expect the Simpson stimulus to lead to increased or decreased levels of cell growth? Please provide your reasoning. B. You construct a series of cell lines with mutations in each one of these genes...