FIG 3.

Immunoreactivity of TESA with Chagasic sera. (A) Western blots containing whole-cell extracts of L. donovani promastigotes (lane 1), T. brucei procyclics (lane 2), T. cruzi trypomastigotes (lane 3), or TESA EVs (lane 4) were probed with pooled sera from Colombian uninfected controls or Chagasic patients with various stages of cardiopathologies as previously detailed (54). (B) IgG antibodies isolated from uninfected controls or Chagas patients with ventricular arrhythmia were coupled to a protein G column, and reactive proteins were immunoprecipitated using these affinity matrices for Western blotting. Proteins in the total TESA EV fraction (lane 1), Triton X-100 extracts of TESA EVs (lane 2), precipitated by uninfected control IgG (lane 3), or IgG from Chagas patients (lane 4) were resolved by SDS-PAGE, and the membranes were probed with biotinylated IgG antibodies from uninfected subjects or Chagas-infected subjects. Primary antibodies were detected using streptavidin-horseradish peroxidase (Western blot membranes were exposed on film for 60 s).