Cells: T293 kidney embryonic cells

Experiment:  
	- over-expression of Musashi-1 (MSI) protein, compared to GFP control (GFP)
	- cells harvested after 24hrs
	- cell homogenized with dounce homogenizer and cytosol (Cytosol fraction) collected after centrifugation
	
To prepare protein extracts, Daoy cells were re-suspended in an equal volume of lysis buffer (50mM Tris pH 8.0, 50mM NaCl, 1mM EDTA and 1 tablet of Complete Proteinase Inhibitor (Roche)/10ml) and incubated on ice for 30 minutes. Cells were then lysed with a dounce homogenizer and the soluble protein extracts obtained after centrifugation.	

	- pellet re-homogenized with dounce homogenizer and via passage through syringe (Pellet fraction)
	- LC-MS/MS analysis on LTQ-Orbitrap of MSI/GFP in Cytosol/Pellet
	
CYS modification: 
YES

Mass spectrometry:
LC-MS/MS on LTQ-Orbitrap

Analysis:
SEQUEST (Bioworks)
Peptide/ProteinProphet v4.0