The images of representive 2D-DIGE and validation of vimentin. (A) A representative 2D-DIGE gel images. The extracted proteins were labeled with fluorescent dyes and separated by 2D-DIGE. B16M group was labled with cy3, B16 group was labled with cy5. (B) A representative two-dimensional gel image. Differential expressed proteins that have been successfully identified by MALDI-TOF/MS (p ≤ 0.05, protein fold≥2) are circled and numbered. The spot numbers correspond to those proteins listed in Table 1. (C) The magnified protein spot images of vimentin in 2D gel showing the significant over-expression in B16M group compared with B16 group. (D) Western blotting shows changes in expression levels of vimentin in B16M group and B16M group; β-actin is used as the internal loading control. (E) Histogram showing the relative expression levels of vimentin in eight pairs of B16M and B16 tissues, as determined by densitometric analysis (p = 0.021).