2D-DIGE analysis of the <i>L.major</i> mock and GFP-PA2G4 proteome.

<p>(A) Representative 2D-DIGE gel. Protein extracts from promastigotes of three independent biological experiments were differentially labeled with the CyDye fluors Cy3, Cy5 and Cy2, and separated by two-dimensional electrophoresis on 13 cm pH 4–7 IPG strips and 12.5% polyacrylamide gels. Gels were scanned on a Typhoon FLA 9500 (GE Healthcare) imager. A merged image of Cy5-labeled GFP-mock (red) and Cy3-labeled GFP-PA2G4 (green) is shown. The molecular weight of marker proteins and the pI range of the IEF gradient are indicated. (B) Differences in protein abundance were revealed by analyzing the gel images with Delta2D v4.3 software (Decodon). Histograms are a graphic representation of these significant differences. For normalization purposes, a Cy2-labeled internal standard was included, corresponding to a pool of protein from all extracts used in the analysis (yellow bars). Fold change and p-values of the spots selected are shown, as well as protein identification by mass spectrometry. Raw data of MS data and Mascot searches is presented in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002646#pntd.0002646.s003" target="_blank">Table S1</a>.