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.