, 2012), we tested whether the infecting T. cruzi strain affects behavioral changes. Lastly, because an immunological dysbalance with high TNF plasma levels is a feature of chronic Chagas this website disease ( Dutra et al., 2009 and Lannes-Vieira et al., 2011), we also investigated the existence of an inflammatory component in T. cruzi-induced depressive-like behavior by targeting TNF. Four- to six-week-old female mice of the C3H/He (H-2k) or C57BL/6 (H-2b) lineages with an average weight of between

15–22 g were obtained from the animal facilities of the Oswaldo Cruz Foundation (CECAL, Rio de Janeiro, Brazil). The infected and uninfected experimental groups of animals consisted of 3–10 mice per group. All experimental procedures were repeated twice or 3 times. The experimental groups consisted of the following: 6 animals of each lineage per experiment to follow parasitemia ( Fig. 1A); 10 animals of each lineage per experiment to follow survival

( Fig. 1B); 5 animals of each lineage per experimental point to follow CNS inflammation and parasitism ( Fig. 1A and B, Table S1); 10 non-infected (NI), 8 acutely and 6 chronically T. cruzi-infected C57BL/6 mice for examination in the open-field test ( Fig. S1); 8 NI and 9 T. cruzi-infected mice of each lineage per experimental point for examination in the open-field test ( Fig. 2); 10 NI, 7 acutely and 7 chronically T. cruzi-infected C3H/He mice for examination in the open-field test ( Fig. S2); 3 NI and 5 T. cruzi-infected C57BL/6 mice and 4 NI and 6 T. cruzi-infected C3H/He mice per experiment to evaluate body weight in a kinetic study ( Fig. S3A and S3B); 7 NI and 8 T. cruzi-infected PLX-4720 cell line C57BL/6 or C3H/He mice per experimental point to evaluate body weight and temperature ( Fig. S3C-S3H); 8 NI and 9 T. cruzi-infected C3H/He mice per experimental point and 5 NI and 10 T. cruzi-infected C57BL/6 mice per experimental point subjected

to the tail-suspension test Ibrutinib (TST) and, sequentially, to the forced-swim test (FST) ( Fig. 3); 6 T. cruzi-infected C3H/He mice to follow parasitemia ( Fig. 4A); 3 NI and 5 T. cruzi-infected C3H/He mice per experimental point to perform TST and immunohistochemical studies ( Fig. 4B–D); 9 NI and 30 T. cruzi-infected C3H/He mice for the kinetic study ( Fig. 5A); 4–5 NI and 7–10 T. cruzi-infected C3H/He or C57BL/6 mice per experimental group to be checked for body weight and rectal temperature, submitted to the indicated treatment and subjected to the TST and, sequentially, to the FST and studies for detection of mRNA ( Fig. 5B–F); 10 NI and 5–16 acutely T. cruzi-infected C3H/He mice per experimental group submitted to the indicated treatment and subjected to the TST ( Fig. 6A and B, Table 1) and studies for detection of mRNA ( Fig. 7C); 5 NI and 3–10 chronically T. cruzi-infected C3H/He mice per experimental group submitted to the indicated treatment and subjected to the TST ( Fig. 6C); 3–4 NI and 4–10 T.

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