<p>Loss of <i>Pf</i>ATG18 causes hemozoin dislocation and parasite death.</p>

<p>Synchronized parasites were treated with DMSO or RAP from the ring stage onward. <b>(A)</b> RAP-induced excision of <i>loxP</i>-flanked genomic DNA in <i>atg18-mNG:loxP</i> parasites, confirmed by diagnostic PCR 5 (see <a href="plospathogens.org/article/info:doi/10.1371/journal.ppat.1014608#ppat.1014608.g001" target="_blank">Figure 1A</a>). <b>(B-E)</b> RAP efficiently depletes <i>Pf</i>ATG18-mNG by 36 hours post invasion, as shown by Western blot (B) and live-cell imaging <b>(C)</b>.

Quantitative fluorescence microscopy reveals strong reduction of <i>Pf</i>ATG18 <b>(D)</b>, but not CRT <b>(E)</b>, following RAP treatment. Parental B11 parasites [<a href="plospathogens.org/article/info:doi/10.1371/journal.ppat.1014608#ppat.1014608.ref043" target="_blank">43</a>] serve as background control. Mean + /- SD; one-way ANOVA with Tukey’s test; n = 99 parasites, 3 experiments. n.s., <i>p</i> > 0.05; ***, <i>p</i> < 0.001. <b>(F-H)</b> <i>Pf</i>ATG18 is essential for asexual blood-stage development.