Fig. 4.
Representative difference gel electrophoresis (DIGE) images of protein extracts of Prdx6−/− and Prdx6+/+ LECs. Labeled samples (Cy3 for Prdx6−/− LECs in green and Cy5 for Prdx6+/+ LECs in red) were subjected to 2-dimensional (2D) gel electrophoresis. Fluorescence was scanned, and the derived images were superimposed with pseudocolors in the dyes. Proteins that are up- or downregulated in Prdx6−/− cells compared with Prdx+/+ LECs are represented by red (up) and green (down) spots, whereas proteins that are equally abundant in both samples appear yellow. The full range of the horizontal axis is from 4 (left) to 7 (right) pH units, and the full range of the vertical axis is from ∼10 (bottom) to ∼100 (top) kDa. A: superimposed images from Cy3- and Cy5-labeled samples in the large gel (13 × 13 cm). Arrows and spot numbers (see Table 1) indicate significant upregulated (red) or downregulated (green) proteins present in Prdx6−/− LECs vs. Prdx6+/+ LECs. B: enlarged area from a small (7 × 7 cm) 2D-SDS gel showing Prdx6−/− (left) and Prdx6+/+ (right) LECs. Protein spots that were decreased or increased >1.5-fold were selected for matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) identification. Mr, molecular weight ratio.