Fig. 9. CP and chagasin expression in different T. cruzi isolates. (A) Immunoblotting of epimastigote lysates probed with monoclonal antibody against cruzipain, monoclonal antibodies against the cathepsin-B-like enzyme of T. cruzi (TcCB) or monoclonal antibodies against calreticulin. (B) Detection of CP activity in epimastigote lysates (1 µg) in 50 mM Na₂HPO₄, 200 mM NaCl, 5 mM EDTA, pH 6.5, 2.5 mM DTT, 5% DMSO at room temperature using 5 µM CBZ-Phe-Arg-AMC as a substrate. Substrate hydrolysis was fully inhibited by E-64 (not shown). The experiments were performed in triplicate and are represented as mean values with standard deviations (SD). The variance was analysed using ANOVA and the asterisks indicate the scores that are statistically significant at P<0.001. (C) Immunoblotting of epimastigote lysates probed with anti-chagasin antibodies or with monoclonal antibodies against calreticulin. The relative intensities of the reactive bands were estimated by scanning densitometry normalized to Dm28c and are indicated below each lane.