STED super-resolution and ultrastructure expansion microscopy reveal detailed organization of microtubules, centriolar plaques and centromeres during schizogony. (A) Dual-color STED nanoscopy images of different schizogony stages of 3D7 parasites expressing tagged nuclear pore protein Nup313-HA_glms, labeled with anti-centrin (green), anti-tubulin (magenta) antibodies and stained with Hoechst (blue). Single slices are shown except for the extended spindle. Quantification of percentage of hemispindles in mononucleated cells with and without centrin signal was performed in 3D7 wild-type cells (n = 52 cells, 1 replicate) imaged with confocal microscopy. (B) Confocal U-ExM images of individual schizont nuclei of the 3D7 Nup313-3xHA_glms strain in hemispindle and mitotic spindle phase, labeled as in (A), except for using three instead of one anti-tubulin antibody. Maximum intensity projections are shown. (C) Quantification of lengths (n = 217, corrected by a measured expansion factor of 4.5) and number of hemispindle branches per nucleus (n = 38) and mitotic spindle lengths (n = 38) of 3D7 Nup313-3xHA_glms expressing cells imaged with U-ExM in 2 replicates. (D) Like (A) with anti-tubulin (magenta) and anti-CenH3 (yellow) showing centromere positioning in hemispindle phases. All scale bars are 1 μm.

Simon CS, Funaya C, Bauer J, Voβ Y, Machado M, Penning A, Klaschka D, Cyrklaff M, Kim J, Ganter M, Guizetti J. An extended DNA-free intranuclear compartment organizes centrosome microtubules in malaria parasites. Life Sci Alliance. 2021 4(11):e202101199.