Fig. 3: Ultrastructure expansion microscopy (U-ExM) detecting SPMT association of DHC3.

A U-ExM of DHC3 in the dhc3::6HA ookinetes. The parasites were stained with the anti-HA antibody. Three independent experiments with similar results. Scale bars: 5 μm. B U-ExM of DHC3 in the 4Myc::dhc3 ookinetes. The parasites were stained with the anti-Myc antibody. Three independent experiments with similar results. Scale bars: 5 μm. C U-ExM of DHC3 (HA) and SPMTs (α- and β-Tubulin) in the early and mature ookinetes of the dhc3::6HA parasites. The parasites were co-stained with the anti-HA antibody and anti-α/β Tubulin antibodies. White asterisk indicates the spindle while the white arrow indicates the apical tubulin ring (ATR). Three independent experiments with similar results. Scale bars: 5 μm. D Diagram showing the microtubule-binding domain (MTBD, green) and truncation of MTBD in DHC3 from the parental parasite dhc3::6HA, generating the modified line ΔMTBD. The green number indicates the structural boundary of MTBD. E Immunoblot of HA-tagged DHC3 in the gametocytes of the dhc3::6HA and ΔMTBD parasites. BiP as a loading control. Two independent experiments with similar results. F In vitro ookinete formation of the dhc3::6HA and ΔMTBD parasites. Values are means ± SEM (n = 3). A two-sided t-test was applied. G IFA of DHC3 expression in ookinetes of the dhc3::6HA and ΔMTBD parasites. Three independent experiments. Scale bars: 5 μm. H U-ExM of DHC3 and SPMTs in early stage (left panel) and later-stage ookinetes (right panel) of the dhc3::6HA and ΔMTBD parasites. Parasites were co-stained with antibodies against HA and Tubulin polyglutamylation (PolyE). PolyE is a marker for the stabilized MT. Three independent experiments with similar results. Scale bars: 5 μm. Source data are provided as a Source Data file.