The detection of ROS level in parasitized-erythrocyte was performed using H2DCFDA dye. For this, parasite culture was treated with compounds for 48 h and then washed with 1X PBS. Treated parasites were incubated with H2DCFDA dye (5 µM) and incubated for 20 min in dark and subsequently washed using 1X PBS. Thin blood smears were prepared on glass slides and mounted using anti-fade DAPI. ROS generation was detected under confocal microscope (Nikon Eclipse Ti2-E) using 100× magnification. Intensity of generated signal was calculated using NIS-Elements AR analysis 5.20.02 software.

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