Gels were stained with silver nitrate at room temperature for 30 min and digital images of protein dots on the gels were captured using a scanner (Seiko Epson Corporation). PDQuest 8.0 software (Bio-Rad Laboratories, Inc.) was used to identify differential spots between the Control, I/R and D-Post groups as previously described (9 (link),25 ), and these differential protein spots were excised from the gels and digested. The peptide mass fingerprint was obtained using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) with a mass spectrometer (Ultraflex III; Bruker Corporation) and compared with that from the NCBInr protein database (http://www.matrixscience.com/help/seq_db_setup_nr.html) as reported previously (9 (link)). Peptides were extracted with 50 mM NH4HCO3:ACN (1:1, v/v). The peptide solution (3 µl) was applied to a target disk to evaporate, and mixed with 0.1 µl matrix solution (4 mg/ml in 70% ACN and 30% 0.1% TFA, v/v), spectra was obtained with MALDI TOF/TOF mass. BioTools 3.0 (Bruker Corporation) and Mascot software (Matrix Science, Inc.) were the databases used to identify proteins via peptide mass fingerprinting. NCBInr was chosen as the sequence database. The names of the differentially expressed proteins were then confirmed.