Fig. 1: PfNDC80-HA and PfNuf2-HA localisation in P. falciparum asexual blood stages.

a Illustration of P. falciparum mitotic stages. The diagrams represent parasites with a hemispindle, a mitotic spindle, an interpolar spindle, a spindle remnant and a formed merozoite. The apical complex is assembled above a nuclear membrane-embedded centriolar plaque, with inner and outer regions. Structures are labelled in the diagrams or defined in the key at the bottom of the diagram. b U-ExM images of schizont-infected RBCs labelled with DyLight488-NHS-ester in BSA (NHS-BSA, greyscale and green) or DyLight488-NHS-ester in PBS (NHS-PBS, greyscale, and inverted greyscale). NHS-BSA (left panels) gives good labelling of membranes. The nuclear membranes are highlighted. NHS-PBS (right panels; with zoom inset) gives good labelling of protein-rich structures, such as the rhoptries and centriolar plaque. U-ExM images of (c) PfNDC80-HA transfectants (anti-HA (PfNDC80), greyscale, yellow arrowheads), counter-stained with NHS-BSA (green) and (d) PfNuf2-HA/GFP-PfCENH3 transfectants (anti-HA (PfNuf2), green, and anti-GFP (PfCENH3); red, yellow arrowheads) counter-stained with NHS-PBS (greyscale), revealing the kinetochore and centromere components co-located and relocating from the base of the hemispindles to the mid-plane of the mitotic spindle, then contracting to the nuclear periphery. The images are displayed as z-projections. Anti-β-tubulin (anti-β-tub, red) marks the spindle microtubules (intranuclear) and sub-pellicular microtubules (SP-MT, white arrowheads). DAPI (blue) stains the chromatin. Zoom-out/zoom-in scale bars are 5 μm and 2 μm. Additional images/ channels are presented in Supplementary Fig. 2 (PfNDC80-HA), Supplementary Fig. 3 (PfNuf2-HA) and Supplementary Fig. 5a (PfNuf2-HA/ GFP-PfCENH3) and 5b (PfNDC80-HA/ GFP- PfCENH3). Figure 1c, d experiments were performed 3 times from 3 different sample preparations.