Figure 1.
Growth of T. brucei procyclic form cells in original SDM-79 and SILAC labelling media.
A. Cumulative growth curve. Growth in original SDM-79 containing non-dialysed FBS (open squares) is shown in parallel to SDM-79+R0K0 (open circles) and SDM-79+R6K6 (closed circles), both containing dialysed FBS. B. DIC light microscopy. T. brucei procyclic cells grown in original SDM-79, SDM-79+R0K0 or SDM-79+R6K6 for ten days were fixed in 4% paraformaldehyde and DIC images acquired on a Zeiss confocal microscope.
Figure 2.
Procyclic cells were cultured in SDM-79+R6K6 then mixed 1∶1 with either unlabeled procyclic or bloodstream form cells. Sample complexity was reduced prior to LC-MS/MS analysis by either fractionation at the protein level by SDS-PAGE or at the peptide level by SCX chromatography.
Figure 3.
Histograms of Log2 fold change.
A. Procyclic form labeled with heavy isotopes (R6K6) mixed 1∶1 with unlabeled procyclic form (R0K0). B. Procyclic form labeled with heavy isotopes (R6K6) mixed 1∶1 with unlabeled bloodstream form (R0K0).
Figure 4.
Agreement of comparative proteomic data with known biology.
Heatmap showing the Log2 FC (procyclic to bloodstream) derived from comparative proteomic data (this study) and previous transcriptomic studies [10], [11], [12]. Grey – not observed. Heatmap generated with GENEE (http://www.broadinstitute.org/cancer/software/GENE-E/).
Figure 5.
Comparison of Proteomic and transcriptomic data.
Scatterplot of the Log2 FC (procyclic to bloodstream) derived from comparative proteomic data (this study) and previous transcriptomic studies [10], [11], [12].
Figure 6.
A. Proteins with greater than ten-fold up- or down regulation, with enrichment P<0.01. B. Constitutively expressed proteins, with enrichment P<0.01.
Table 1.
Negative correlation between protein and Mrna.