Fig. 4

Depletion of PPKL caused defects in daughter parasite maturation and parasite morphology. A, B Parasite morphology was strongly affected by depletion of PPKL. Parasites were grown in the presence or absence of auxin for 24 h, followed by fixation for IFA with antibodies against actin, tubulin, and acetylated tubulin. IMC1 and GAP45 were used as controls for the IFA. Three independent experiments were performed, with similar outcomes. Scale bars = 2 μm. C–E The maturation of the daughter parasites and cytoskeleton integrity were strongly disrupted upon depletion of PPKL. Parasites were grown for 24 h in total, and the PPKL-AID line were induced in auxin for 6, 12, and 24 h (h). The parasites were examined by IFA using antibodies against GAP45, IMC1, tubulin, and centrin1. The parasite body was enlarged with clear emerging daughter parasites, multiple nuclei, and elongated cytoskeleton fibers in PPKL-depleted parasites. Scale = 5 μm. F Quantification of parasites stained with GAP45 and tubulin. The parasites observed in (C–E) were scored for the calculation of parasites with normal morphology (black column), parasites with enlarged bodies (stained by GAP45, red column), and parasites with elongated cytoskeleton fibers (stained by tubulin, red column)