Figure 4.

Experimental design, coagulation test aPTT and tail bleeding assay in NI+V, OI+V, OI+isotype, NI+T and OI+T mice. (A) Male BALB/c mice were infected with 5 × 10⁴ insect-derived metacyclic forms of T. cruzi (Tulahuén strain) within the oral cavity. Anti-IL6R or IgG treatment began after 14 dpi and was performed in each 48 h. (B) Bleeding was caused by a tail transection in NI+Vehicle (NI+V), OI+V, OI+isotype, NI+treatment (NI+T) and OI+T. Absorbance at 540 nm (hemoglobin concentration) was used to estimate blood loss. (C) NI+V, OI+V, OI+isotype, NI+T and OI+T plasma were obtained by cardiac puncture followed by addition of the aPTT reagent as described in the “Methods” section. Clotting time was estimated using a coagulometer. (B,C) Values are presented as mean ± SEM for each group/day post-infection and are representative of three independent experiments. Results were analyzed using one way ANOVA with Tukey's multiple comparisons test (*0.0001< p < 0.05; #p < 0.0001).