*2.1. E. histolytica Trophozoites*

*E. histolytica* trophozoites (HM1:IMSS strain) were axenically cultured in TYI-S-33 medium supplemented with Diamond vitamin tween solution (Merck) and 15% heatinactivated adult bovine serum (Microlab). Trophozoites were grown for 72 h at 37 ◦C until they reached the log phase and harvested by chilling on ice for 5 min and centrifugation at 1400 rpm during 5 min at 10 ◦C. The pellet was resuspended in PBS pH 7.4 and the trophozoites counted in hemocytometer and preserved at room temperature until use. For selected experiments, amoebas (5 × 105) were resuspended in 500 <sup>μ</sup>L of PBS and fixed with formaldehyde (3.7%) or heat-inactivated (56 ◦C) for 30 min in both cases.
