Fig. 4: PfNDC80-deficient parasites fail to form intact merozoites.

a, b Representative U-ExM images of segmented schizonts in control (DMSO) and PfNDC80-cKO/KD (conditional knockout/knockdown, treated with rapamycin and glucosamine) parasites. The sub-pellicular microtubules are labelled with anti-β-tubulin (anti-β-tub, red, aqua arrowheads) and the chromatin with DAPI (blue). NHS-BSA (green) counterstains the membranes. a Anti-PfGAP45 labels the inner membrane complex (anti-GAP45, greyscale, magenta arrowheads). The experiments were performed twice. b Anti-polyE labels the sub-pellicular microtubules at their minus ends (greyscale, aqua arrowheads). The experiment was performed one time. All images are presented as z-projections. All scale bars are 5 μm, except the zoom images which are 2 μm. Additional images are presented in Supplementary Fig. 12a, b. c, d. Quantification of nuclei parameters for control and PfNDC80-cKO/KD segmented schizonts based on U-ExM data (n = 24 cells). The fraction of unpackaged nuclei c and akaryotic merozoites d are presented (n = 24 cells). Statistical differences were determined using an unpaired Welch’s t-test for c and d (****p < 0.0001). Further quantitative analysis is presented in Supplementary Fig. 12c–f. Figure 4c, d images for analysis were acquired from three independent experiments. e Live cell imaging of control and PfNDC80-cKO/KD segmented schizonts before (upper row) and after (lower row) egress. The subpellicular microtubules were labelled with Tubulin Tracker (TT, red) and chromatin was labelled with Hoechst (greyscale). The yellow arrowheads indicate akaryotic merozoites and blue arrowheads indicate naked nuclei. All images are presented as z-projections. The scale bar is 2 μm. The experiments were performed 3 times. f Single section electron micrographs of segmented schizonts in control and PfNDC80-cKO/KD parasites. The nuclei (N) are marked and the rhoptries are indicated with yellow arrowheads. g 3-D array tomography reconstructions of segmented schizonts in control and PfNDC80-cKO/KD parasites. Figure 5f, g experiments were performed one time. Colour legend: Green, nucleated daughter merozoite plasma membrane; red, anucleate daughter merozoite plasma membrane; blue, nuclei; yellow, rhoptries. f Scale bar is 1 μm.