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Introduction
2D DIGE image overlays enable easy visual comparison between two protein samples labeled with Cy3 and Cy5 and presented in green and red colors. The more mature B cell type is presented in red color. Triple image overlays enable comparison between the two samples in relative to internal standard sample (Stnd) labeled with Cy2 and presented in blue color.
Explanation for colors in double image overlays:
Yellow: Proteins shared between the two samples.
Green and red: Unique proteins in the sample. In Ramos samples (Gels 5-7) red indicates up-regulation and green down-regulation in response to anti-IgM stimulation.
Triple image overlays:
White: Proteins shared between the three samples (no difference in protein abundance).
Yellow: Proteins shared between the two samples. Protein abundance is high in the two samples in relative to Stnd.
Blue: Protein abundance is negligible in relative to Stnd.
Green and red: Unique proteins in the sample. Protein abundance is high in relative to Stnd and to the other sample.
Turquoise and purple: Protein abundance is higher/lower in relative to the other sample.
Highlighted
Proteins highlighted in red are identified either by PMF or by combining MS and MS/MS spectral data for database searches. Proteins highlighted in blue are identified by comparing their positions on 2D DIGE gel to known proteins in other lymphocyte 2DE databases. Move the mouse pointer over a spot to see the protein name and number referring to Table 1 and Supplementary Tables 2-3. Click on the spot to get UniProt information about the protein.
Original view
Interactive 2D DIGE map. B cell protein samples labeled with Cy3 and Cy5 and separated in nonlinear pH gradient 3-10. Internal standard is labeled with Cy2 and run on the same gel.
© 2012
University of Tampere
IBT Bioinformatics
Finland
Lund University
Department of Experimental Medical Science
Sweden
Interactive 2D DIGE maps
Proteome samples from 24 h anti-IgM stimulated Ramos cells and corresponding control cells are labeled with Cy5 and Cy3 dyes, respectively, and separated in nonlinear pH gradient 3-10. Internal standard (Stnd) is pooled reference sample labeled with Cy2 and run on the same gel. Move the mouse pointer over a spot to see the protein name and number referring to Table 1 and Supplementary Tables 2-3. Click on the spot to get UniProt information about the protein.
Middle events in Ramos B cell activation (up- and down-regulated proteins in colors):
Double image overlay highlights the 24 h anti-IgM stimulation induced changes in Ramos proteome. Red proteins are up-regulated and green are down-regulated in response to anti-IgM stimulation. Yellow proteins are shared between the samples (no change).
Triple image overlay enable comparison between 24 h stimulated (Cy5 red) and 24 h control Ramos (Cy3 green) in relative to Stnd (Cy2 blue). Red and purple proteins are up-regulated, whereas green and turquoise proteins are down-regulated. Yellow color indicates that the relative protein abundance is high in Ramos cells but does not change in response to stimulation. Blue color indicates low protein abundance in Ramos cells compared to Stnd. White proteins are shared between the three samples (no difference).