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Figure 3 | BMC Microbiology

Figure 3

From: The Trypanosoma cruzi nucleic acid binding protein Tc38 presents changes in the intramitochondrial distribution during the cell cycle

Figure 3

Subcellular localization of Tc38 using biochemical approaches in T. cruzi epimastigotes. (A) Digitonin extraction. Epimastigotes (125 mg per tube) were incubated with different digitonin concentrations (indicated on the abscissa) as described in Materials and Methods. Marker enzymes activities: hexokinase (HK), citrate synthase (CS), and pyruvate kinase (PK). The amounts of Tc38 were determined by western analysis. (B) Subcellular fractionation. The experiment was carried out using 3.3 g (wet weight) of parasites. Fractions are plotted in the order of their isolation, from left to right: nuclear (N), large granule (G), small granule (SG), microsomal (M) and supernatant (S). The ordinate represents relative specific activity (percentage of total activity/percentage of total protein). The abscissa indicates the cumulative protein content. The percentage of recovery for the marker enzymes: citrate synthase 70.9%, hexokinase 74.1%, cytochrome C reductase 43.6%, pyruvate kinase 85.3%, Tc38 61.1%. Error bars indicate the variation in band intensity seen by quantification of the western blot.

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