Identification of selected protein spots extracted from 2D-SDS-PAGE.

Protein spots were extracted from 2D- polyacrylamide gel with an Eppendorf pipette and approx. 1 mm in diameter gel pieces were destained according to the procedure described by Gharahdaghi et al. [42]. In-gel tryptic digestion of proteins was done using Promega (Madison, WI, USA) Gold Label trypsin, following the manufacturer’s protocol with only minor alterations. The tryptic digest sample (1–10 µl) was injected on a trap column coupled to a C18 Waters NanoEase capillary column 75 µm×50 mm. The ESI-MS/MS experiments were carried out on a QTOF-Global mass spectrometer (Micromass, Manchester, UK).*Mascot score was obtained after submitting raw data (PKL format) to the Matrix Science website for evaluation (http://www.matrixscience.com/cgi/search_form.pl?FORMVER=2&SEARCH=MIS).