*2.14. Detection of ROS*

WT trophozoites, AFAT, and WT trophozoites that were cultivated with AF (2 μM) for 24 h (WT + AF acute) were incubated with 0.4 mM H2DCFDA for 15 min at 37 ◦C. The trophozoites were washed twice with PBS, and the level of oxidation was analyzed by flow cytometry. Flow cytometry was performed using Cyan ADP flow cytometer (Agilent Dako, CA, USA) and data from 10,000 cells were collected for each condition.

#### *2.15. Detection of OXs by RAC (OX-RAC)*

The detection of OXs by OX-RAC was performed using a previously described protocol [31]. A protein was considered to be oxidized when its relative amount in the DTTtreated lysates was at least two times greater than that in the untreated lysates (*p* < 0.05 according to the results of an unpaired *t*-test).
