Global transcriptomic changes of Plasmodium falciparum asexual parasites and late gametocytes after treatment with MMV390048 or MMV642943

Plasmodium falciparum asexual parasites and late gametocytes were treated for 24 or 48 h with MMV390048 or MMV642943 in order to understand the functional consequences of inhibiting lipid kinases Agilent Plasmodium falciparum DNA microarrays were performed to analyze the global transcriptomic profiles of asexual parasites and late stage gametocytes following 24 or 48 h treatment with MMV390048 or MMV642943. NF54 ring-stage ( >95% synchronicity, 6 hours post merozoite invasion) asexual parasites and mixed stage IV and V gametocytes were treated with 10 X IC50 of MMV390048 or MMV642943 for 24 or 48 h. Thereafter, parasitised erythrocytes were washed and stored at -70°C. Gametocytes were isolated from erythrocytes using Nycoprep, washed and stored at -70°C. RNA was isolated using a a combination of TriZol reagent and Qiagen RNeasy kit. RNA from each of the 10 samples labelled with Cy5 was hybridised against Cy3 labelled reference pool consisting of untreated and treated mixed asexual parasites and gametocytes. Arrays were scanned using the Axon GenePix 4000B scanner. Images were quantified and data was normalized with GenePix Pro 5.1 software.