Cells: human embryonic kidney cells T293

Experiment:  

The T293 cell line was obtained from American Type Culture Collection (ATCC). Cells are over-expressing GFP in lentivirus construct. 
Cells were cultured in improved minimum essential medium (IMEM) (Invitrogen, Carlsbad, CA) 
supplemented with 10% fetal bovine serum (Atlanta Biologicals, Inc., Lawrenceville, GA). Cells were grown until 90% confluence and then harvested to prepare RNA and protein extracts for further 
analyses.

To prepare protein extracts, 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. 
To obtain the 'pellet' fraction (labeled NUCL), the pellet was re-homogenized twice and soluble fraction extracted after centrifugation. 

CYS modification: 
YES

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

Analysis:

SEQUEST (Bioworks)
Peptide/ProteinProphet v4.0