Fig. 2.
Effect of divalent metal chelators and anti-Tcgp63-I antibodies on T. cruzi–R. prolixus interaction. Dm28c epimastigotes (2 × 107 cells) were treated for 1 h at 27 °C with 1 µm EGTA, EDTA or phenanthroline at 0.2, 0.3 or 0.5 µm (A). Alternatively, the parasites were treated for 60 min at room temperature with anti-Tcgp63-I antibodies at 1:500 and 1:1000 dilution, or pre-immune serum at 1:500 (B). Parasite viability was not affected by the treatments used in this set of experiments. Following interaction for 15 min with the insect gut, the number of adhered parasites/insect gut epithelial cells was estimated by randomly counting at least 100 epithelial cells in quadruplicate in 10 random fields. The results are shown as the mean ± s.e.m. of two independent experiments. An asterisk denotes significantly different from control (untreated cells) using Student's t test (*P < 0.001). Lastly, western blotting analyses identified two reactive bands against T. cruzi Dm28c epimastigotes (a), while no reactive molecules were recognized by the pre-immune serum (b) (C).