β-Hematin inhibition, parasite culturing and cellular heme measurements

The detergent mediated assay for β-hematin inhibition was based on that developed by Carter et al.³⁰ modified to use pyridine to detect unconverted hematin as previously described,³¹ with modifications reported in detail elsewhere.³²

Parasites were cultured according to the method of Trager and Jensen with minor modifications.³³ Parasite growth inhibition experiments were carried out using previously reported methods. For most experiments the parasite lactate dehydrogenase assay was used to measure parasite growth,³⁴ but for the experiments investigating speed of killing and the effects of NAC and Luperox® a SYBR Green I assay was used.^35–36

Measurement of cellular hemoglobin, exchangeable heme and hemozoin was performed as previously described in detail⁸.