Global transcriptomic changes in P. falciparum NF54 gametocytes treated with the histone demethylase inhibitor ML324

Global transcriptomic profiling of ML324 treated and untreated control gametocytes to determine the downstream effects of histone methyltransferase inhibition on gametocyte development Gene expression profiling with microarrays (Agilent Technologies, USA, (Painter, H. J. Methods in Molecular Biology 923: 213-219, doi:10.1007/978-1-62703-26-7_14 (2013)) was performed to determine the global transcriptomic changes in P. falciparum gametocytes arising from treatment with ML324. P. falciparum NF54 gametocytes were induced by culturing parasites in glucose-deficient media with a concurrent drop in haematocrit as described previously (Reader, J. et al. Malaria Journal 14, 213, doi:10.1186/s12936-015-0718-z (2015)). Gametocytes (1-3% gametocytaemia, 4% haematocrit) were treated with ML324 (5 µM) and sampled 24 h thereafter using 0.01% (w/v) saponin. RNA was extracted and cDNA synthesised and dye-coupled with Cy5 as previously described (Painter, H. J. Methods in Molecular Biology 923: 213-219, doi:10.1007/978-1-62703-26-7_14 (2013)). The ML324 treated and paired untreated sample were hybridised to arrays with an equal quantity of Cy3-labeled reference pool cDNA that consisted of a 3D7 mixed asexual parasite population and the untreated and treated gametocyte samples. Slides were scanned with an Agilent G2600D scanner using methods previously described (Painter, H. J. Methods in Molecular Biology 923: 213-219, doi:10.1007/978-1-62703-26-7_14 (2013)).