<p><i>Pf</i>ATG18 is critical for V-ATPase integrity.</p>

Analysis as in <b>(B,C)</b>. <b>(F)</b> V₁B dissociation precedes DV fragmentation. Loss of <i>Pf</i>ATG18 was induced at 28 hpi, shifting the onset of DV fragmentation to 36 hpi. Analysis as in <b>(C)</b>.

Individual and mean values; one-way ANOVA with Tukey’s test; n = 165 parasites, 5 experiments. n.s., <i>p</i> > 0.05; **, <i>p</i> < 0.01. <b>(G)</b> Conditional inactivation of subunit V_oc recapitulates the <i>Pf</i>ATG18 knockout phenotype, with DV fission and HTC accumulation (arrowheads). Shown are CRT-mNG and the first 180 amino acids of glycophorin-binding protein 130 fused to mSc (GBP130_1-180-mSc), an exported and reinternalized marker.

(H,I) Co-immunoprecipitation combined with mass spectrometry identifies candidate <i>Pf</i>ATG18 interaction partners involved in phosphoinositide balance and membrane trafficking. <b>(H)</b> Protein hits ranked by enrichment score, defined as the log₂ fold-change (<i>atg18-mNG:loxP</i>/ B11) multiplied by the negative log₁₀ of the p-value. Benjamini-Hochberg corrected <i>t</i>-test; n = 3 experimen<i>t</i>s with 3 technical replicates each.

Relevant proteins as well as unrelated DVM markers are indicated. <b>(I)</b> Membrane trafficking-related proteins among the top 200 hits with a confirmed or potential role in the endolysosomal pathway. Scale bars: 5 µm. Source data are provided as a <a href="plospathogens.org/article/info:doi/10.1371/journal.ppat.1014608#ppat.1014608.s015" target="_blank">S3 File</a>.</p>