PfAAP6, PfAAP4 and PfAAP2 define a novel, apical structure in P. falciparum. (a) PfAAP6 (PF3D7_0508900), (b) PfAAP4 (PF3D7_1318700) and (c) PfAAP2 (PF3D7_1312800): Schematic representation of each protein (protein length indicated as number of aminoacids) on top. Localization of AAP-GFP fusion proteins in schizonts and free merozoites and PCR-based confirmation of the correct insertion of GFP-encoding integration plasmid into the targeted loci shown underneath. Ladder size indicated in base pairs (bp). gDNA from parental 3D7 was used as control. Nuclei stained with Hoechst-33,342. Scale bars 2μm. (d) Colocalization of PfAAP6-GFP with the rhoptry bulb marker protein ARO-mCherry, the microneme marker AMA1-mCherry and the rhoptry neck marker AIP-mCherry. (e) Colocalization of PfAAP4-GFP with the rhoptry bulb marker protein ARO-mCherry, the microneme marker AMA1-mCherry, the rhoptry neck marker RON12-mCherry, the dense granules marker EXP2-mCherry and the IMC marker ALV5-mCherry. (f) Structured illumination microscopy (SIM) images ofPfAAP4-GFP with the IMC marker ALV5-mCherry during IMC formation. Time points (T) indicated are T1–T3: early schizonts, T4–T6: late schizonts, and T7: free merozoites. Zoom factor: 400%. Scale bars 2μm.

Wichers JS, Wunderlich J, Heincke D, Pazicky S, Strauss J, Schmitt M, Kimmel J, Wilcke L, Scharf S, von Thien H, Burda PC, Spielmann T, Löw C, Filarsky M, Bachmann A, Gilberger TW. Identification of novel inner membrane complex and apical annuli proteins of the malaria parasite Plasmodium falciparum. Cell Microbiol. 2021 23(9):e13341.