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| Figure 1: 2-D gel analysis of differentially expressed proteins in hepatic endosomes obtained from ApoE-/-/B48/48 and wild-type mice. Proteins obtained from mouse liver early or late endosomes were labeled with either Cy3 or Cy5 and subjected to 2-D gel electrophoresis with Cy2-labeled pooled standards. Panels A, B, C, and D show representative 2-D gel images of endosomal proteins obtained from ApoE-/-/B48/48 and wild-type (WT) mice. The spot image was mapped at pI ranged from 4 to 7 NL (left to right) and MW ranged from 100 to 10 kDa (top to bottom). Protein-spot maps from all the gel images were matched and the fluorescence spot intensity was compared between endosomes, as detailed in the Materials and Methods. Two representative protein spots, which are differentially expressed among endosomes, are boxed and circled, respectively. The 3-D images show fluorescence distribution of these spots. The 3-D images labeled with boxed or circled letters correspond to the boxed or circled spots in panels A, B, C, and D. Mass spectrometric analysis demonstrates the boxed and the circled spots as PDIA3 and cathepsin B, as shown in Table 1 (spots 11 and 35). |
