96 well white flat bottom opaque tissue culture plate

GFP-Luciferase expressing Plasmodium falciparum blood stage parasites were cultured asexually. Assays were performed on synchronous (>90%) ring stage parasites at 2% parasitemia and 5% hematocrit. In triplicate, kinase inhibitors were administered to parasites at 500 nM with vehicle as a non-treatment control (0.5% DMSO in PBS), and Chloroquine administered at 2.5 μM as a positive control. At 44 h post treatment, 30 μL of the assay culture was transferred to a 96-well white flat-bottom opaque tissue culture plate (Beckton Dickinson, Franklin Lakes, New Jersey). Using a Berthold LB960 XS3 microplate (Berthold Technologies, Wildbad, Germany) an equal volume of Bright-Glo luciferase reagent (Promega, Madison, WI) was added to parasites and resultant luminescence was measured. Experimental data is representative of three independent experiments.

GFP-Luciferase expressing Plasmodium falciparum blood stage parasites [21 (link)] were cultured asexually. Assays were performed on synchronous (>90%) ring stage parasites at 2% parasitemia and 5% hematocrit. In triplicate, Erastin was administered to parasites at different titrations with vehicle as a non-treatment control (1% DMSO in complete media), and Chloroquine administered at 2.5 µM as a positive control. At 44 h post treatment, 30 µL of the assay culture was transferred to 96-well white flat-bottom opaque tissue culture plate (Beckton Dickinson, Franklin Lakes, NJ). Using a Berthold LB960 XS3 microplate (Berthold Technologies, Wildbad, Germany) an equal volume of Bright-Glo luciferase reagent (Promega, Madison, WI) was added to parasites and resultant luminescence was measured. Experimental data is representative of three biological replicates.