We used live cell imaging to detect expression of each protein throughout the life cycle and to examine their location. The Class XIV myosins were detected predominantly in the invasive stages (developing merozoites in schizonts and merozoites, ookinetes, and developing sporozoites within oocysts and sporozoites). As shown previously, MyoA‐GFP was associated with the surface pellicle of each invasive stage, whereas MyoB‐GFP was localised as an apical end dot in merozoites, mature (>20 hr) ookinetes, and late stage sporozoites. MyoE‐GFP was detected as a dot at the basal end of ookinetes and sporozoites and as a dot in merozoites. MyoE‐GFP was also expressed in liver stages, but as with merozoites, it was not possible to clearly identify the basal end of these stages. MyoF‐GFP was most abundant in the insect stages, particularly ookinetes and oocysts. In mature ookinetes, MyoF‐GFP was restricted to the apical end, whereas in oocysts, it was more evenly distributed throughout the developing sporozoites. In contrast, MyoJ‐GFP was only observed in mature oocysts, located at the junction between the differentiating sporozoites and the oocyst bodies. Following oocyst rupture, MyoJ‐GFP remained associated with the oocyst body and was not present in sporozoites. Finally, MyoK‐GFP was found exclusively associated with the nucleus of gametocytes, appearing as an arc across the cell, and in zygotes/early ookinetes, as two distinctive dots. These dots were either absent or only remnants were seen in mature ookinetes. 

Analyses of mutants expressing tagged myosins provide evidence for the following:
- distinct spatio‐temporal profiles for each myosin during ookinete development

- Colocalisation of myosins with MyoA and the apical marker, ISP1 (PBANKA_1209400; inner membrane complex sub-compartment protein 1)