P. falciparum parasites from routine cultures were seeded as rings at 0.5% initial parasitemia in 1% haematocrit in ACM in triplicate in 96-well plates. Parasites were maintained for 96 h under standard culture conditions and the media was changed daily. At 96 h, parasite cultures were split back to 0.5% parasitemia and maintained as described for an additional 96 h (Supplementary Fig. 1). Parasites were stained at all 0 and 96 h time points with 1 × DNA dye SYBR Green I (Invitrogen) as described in ref. 38 (link), and fixed in 1% paraformaldehyde and 0.0075% glutaraldehyde (Electron Microscopy Sciences) in Alsever's Solution (Sigma-Aldrich) for 30 min at 4 °C. Fixative was removed and cells were stored in PBS at 4 °C until analysis by flow cytometry. Growth rate reflects the fold increase in RBC parasitization between 0 and 96 h. To identify parasitized reticulocytes, cultures were stained with 0.5 μM DNA dye SYTO 61 (Invitrogen)13 (link),39 (link) and PE-conjugated mouse anti-human CD71 antibody (Miltenyi Biotech) and analysed by flow cytometry.