Aberrant motility of <i>P</i>. <i>berghei</i> sporozoites expressing <i>P</i>. <i>falciparum</i> ‘AAA profilin’.

<p>(A) Representative images of ookinetes from the indicated lines moving for 10 minutes. White lines indicate tracks after 10 min. (B) Average speeds of ookinetes with median (line) and mean (arrowheads) from different parasite lines. At least 40 parasites per line were tracked over 15 min; significance was tested with a one-way ANOVA; ** denotes p<0.01, *** p<0.001. Horizontal bar: median, arrowheads indicate mean. (C) 10 ookinetes with median average speeds were plotted for their instantaneous speeds (time lapse: 20 s); significance was tested with a Kruskal-Wallis test; *** denotes p<0.001. Horizontal bars: median, arrowheads: mean. (D-F) Assembly of 40 randomly selected sporozoite tracks with representative time-lapse images below expressing (D) wild type, (E) QNQ and (F) AAA <i>P</i>. <i>falciparum</i> profilin. Images below are 3 seconds apart. Scale bar: 10 μm. Note the different patterns of migration in the AAA mutant. (G) Movement patterns of sporozoites clustered within three groups: ‘persistently moving’ denotes parasites that showed gliding motility for at least 50 consecutive frames (recorded at 0.33 Hz) without their speed dropping below 0.2 μm/s for more then 10 frames; ‘partially moving‘ denotes parasites that showed some circular gliding motility but did not fulfill the ‘persistently moving‘ criteria and ‘attached/waving‘ [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006412#ppat.1006412.ref074" target="_blank">74</a>] denotes parasites that did not display any active circular gliding. (H) Average speeds of persistently moving sporozoites tracked between 50 and 100 frames (150–300 s) for at least 50 sporozoites, *** denotes p<0.001; Kruskal-Wallis test. Horizontal bars: median, arrowheads: mean. (I) 10 sporozoites with median average speeds were plotted for their instantaneous speeds (from one frame to the next = 3 s). Note the shift of AAA sporozoites to a high abundance of very slow speeds as well as some notably high speeds, *** denotes p<0.001; Kruskal-Wallis test. Horizontal bars: median, arrowheads: mean.