Liquid chromatography-mass spectroscopic analysis (LC-MS/MS)
Protein spots separated by 2DE were digested in-gel with trypsin and the
resultant peptides analysed by LC-MS/MS as described by Parker et al.
(2005) and Senko et al. (2003). The LC-MS/MS data were processed with
XCalibur 3.0.63 software (Thermo Fisher Scientific Inc.) and analysed
with the Proteome Discoverer 1.4.0.228 software suite (Thermo Fisher
Scientific Inc.). Statistical analysis of MS data was conducted with
Scaffold Q+S (Proteome Software, Inc.). The abundance of a particular
protein was expressed based on the percentage of Normalized Total
Spectra (NTS) of a specific peptide found in an individual protein spot.