Global transcriptional response of P. falciparum NF54 early gametocytes treated with the Jumonji histone demethylase inhibitor JIB-04

Global transcriptomic profiling of JIB-04 treated and untreated control early gametocytes to determine the downstream effects of Jumonji histone demethylase inhibition on development DNA microarrays (Agilent Technologies, USA, (Painter et al., 2013, DOI: 10.1007/978-1-62703-26-7_14) were used to profile global the transcriptomic changes occuring in P. falciparum gametocytes treated with the Jumonji demethylase inhibitor JIB-04. P. falciparum NF54 gametocytes (1-3% gametocytaemia and 4% haematocrit induced from asexual parasite cultures through a combination glucose starvation and a drop in haematocrit as described previously (Reader et al., 2015, DOI: 10.1186/s12936-1015-0718-z) were treated with 5 µM JIB-04 (Cayman Chemical) on day 2 and day 3 of development and sampled 24 h later with 0.01% (w/v) saponin. RNA was extracted and cDNA synthesised and dye-coupled to Cy5 as previously described (Painter et al., 2013, DOI: 10.1007/978-1-62703-26-7_14). The two treated (Day2_JIB-04 and Day3_JIB-04) and paired untreated samples (Day2_UT and Day3_UT) were each hybridised to an array with an equal quantitiy of Cy3-labeled reference pool cDNA (consisting of untreated and treated NF54 gametocyte samples and mixed 3D7 asexual parasites). Slides were scanned with an Agilent G2600D scanner using methods previously described (Painter et al., 2013, DOI: 10.1007/978-1-62703-26-7_14).