Modulation of the glycerol and glucose metabolic pathways in BSF grown in CMM_Glyc/GlcNAc.

The expression of GK, TAO, ISG75 and FBPase in BSF grown in CMM_Glc and CMM_Glyc/GlcNAc, as well as FBPase in procyclic trypanosomes grown glucose-rich and glucose-depleted conditions was analysed by western blotting using the anti-hsp60 immune serum as control (Panel A). The arrow indicates a 55 kDa band corresponding to GK (calculated Mr: 56.4 kDa), above a non-specific signal produced by the anti-GK immune serum. To compare expression of FBPase/hsp60 the analysis of the BSF and PCF samples was performed on the same gel. Panel B shows the GK and HK enzymatic activities in the three growth conditions (error bars indicate means ± SD of 3 biological replicates). The proteomic data described in S3 Table is presented in Panel C by expressing the percentage of up- or down-regulation of the indicated enzymes in cells grown in CMM_Glyc/GlcNAc as compared to cells grown in CMM_Glc. The dashed line corresponds to two-fold up- or down-regulations. The numbers indicate the enzymatic steps described in Fig 1. The subunits of the pyruvate dehydrogenase complex and of the succinyl-CoA synthetase are annotated 18α-18δ and 20α-20β, respectively. Up- and down-expression of TAO and GK, respectively, are represented by grey bars, and that of glycolytic enzymes are shown with white bars. The asterisks mean that the proteome data have been confirmed by western blotting and/or enzymatic activity assay.