High-throughput Malaria Compound Screening

Small volume 384-well plates (Greiner part 784261) were spotted with 5 μL of each test and control compound, resulting in a 1000× concentration source plate for a 40 nL pin tool (Fig. S1A). For SP assays, libraries were provided preplated at 10 mM in DMSO (MMV). Control compounds primaquine and monensin were plated at 10 mM, MMV390048 (PI4Ki) and atovaquone were plated at 3.3 mM, and nigericin was plated at 200 μM. For DR assays, serial dilutions were made in DMSO (100%) using a Biomek 4000 (Beckman Coulter). Two DR plate maps were used in this study, termed v1 as originally published^{19 (link)} and v2 (Fig. S1B). The assay steps were unchanged regardless of the map version used. Source plates were sealed and shipped on dry ice to Shoklo Malaria Research Unit in Mae Sot, Thailand or Institute Pasteur in Phnom Penh, Cambodia.