Figure 2.

P. falciparum life cycle synchronisation by DFMO. (a) Giemsa-stained morphological evaluation of thin blood smears of DFMO treated parasites for an entire 48 h IDC of P. falciparum 3D7 parasites (5% haematocrit and 3% parasitaemia,1000X enlarged, hpi = hours post-infection). (b) Synchronicity was evaluated through age-binning of parasites based on 2 h evaluation of Giemsa stained parasite morphology for parasites synchronised either with 3 consecutive but 6-hourly spaced sorbitol treatments (3xsorbitol) inspected directly after the last sorbitol treatment, or a single DFMO treatment (IC₉₀) at the next invasion cycle, monitored for a total of 24 h after treatment (C = control, 5% haematocrit and 5% parasitaemia). A minimum of 100 parasites was inspected for each condition. *P < 0.05; ***P < 0.001 calculated using a student t-test. Box plots indicate 80% distribution of data in box and hinges, and 95% in whiskers (c) Quantification of DFMO synchronisation. Percentage of the parasites with increasing DNA content (N = DNA copy number) measured by flow cytometry through SYBR Green I fluorescence collected in the FL-1 channel (FITC signal) on a Becton Dickenson FACSAria with 50 000 total events captured. Parasites were treated with DFMO (IC₉₀) and sampled every 6 h for an entire IDC (48 h). Data are averaged ± S.E. from at least 2 independent biological replicates, performed in technical triplicates.