Fig. 1

Drug treatment strategies used in this study. The lifecycle stage of drug treatment is represented in the first box for each panel and the stage of parasitaemia measurements are highlighted by red boxes with yellow background. (a) Merozoite; successful invasion of drug-treated purified merozoites was measured at ring-stage immediately after addition of erythrocytes and merozoite invasion (<1 hour rings) or after washing out the drug (denoted by green dashed line) and growing parasites through to late trophozoite stage (40 hours post-invasion). (b) In cycle; early ring-stage parasites (<4 hours post-invasion) were drug-treated and the resulting growth inhibition was assessed at late trophozoite stage (40 hours post-invasion). (c) 1 cycle; early ring-stage parasites (<4 hours post-invasion) were drug-treated and the resulting growth inhibition was assessed 90 hours later, after 1 cycle of reinvasion, at late trophozoite stage. These assays are a longer duration, but nonetheless equivalent in terms of including 1 cycle of reinvasion and development, to 1 cycle assays reported in other studies [41–44]. (d) 2 cycle (delayed death); early ring-stage parasites (<4 hours post-invasion) were drug-treated and the drug-treated parasites were grown for 80 hours prior to washing out the drug in fresh media (denoted by green dashed line). Growth inhibition was assessed approximately 40 hours later after a second cycle of reinvasion (120 hours post-invasion). (e) Live filming; mature schizont-stage parasites were incubated with azithromycin and the success of merozoite invasion was recorded and quantified by live filming (green arrow, successful invasion; blue arrow, deformation of the erythrocyte but no invasion; red arrow, attachment and release or attachment with failure to deform erythrocyte and no release) (see Additional file 1: Video S1 and Additional file 2: Video S2). White box-drug treatment; yellow box-analysis of parasitaemia; green dashed line-drug washout