Platelet proteome analysis from control and exposed rats. Platelets were isolated from both control and hypoxia-exposed animals, and total platelet protein was prepared for platelet proteome analysis. Each protein lysate was subjected first to isoelectric focusing, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension, as described in the supplemental Methods. The differentially expressed proteins were identified using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. (A) Representative 2-dimensional gel electrophoresis (2DE) gel image of platelet proteome (4-7 pI range, 13 cm). (B) The pie chart of platelet protein features from 2DE gel after differential analysis of gel images by Progenesis SameSpots software shows that 27% of platelet protein features in range of pH 4 to 7 were altered in hypoxia-exposed platelets. (C) A typical heat map of a selected portion from 2DE gels showing representative differential spots in 3 dimensions, prepared using ImageJ software. (D-E) Bioinformatic analysis of identified proteins was performed with the GeneCodis Web tool (http://genecodis.cnb.csic.es). (D) Gene Ontology cellular compartment analysis and (E) Gene Ontology molecular function analysis. See the supplemental Methods for details.