Fig 7. PfCen1 phase separation depends on its N-terminus and calcium binding.

(A-B) Turbidity measurements in recombinant protein solutions at 30 μM of PfCen1 mutants lacking the N-terminus or with disabled EFh domains, respectively, during addition of calcium followed by EDTA. Conditions: 50 mM BisTris (pH 7.1) at 37°C. (C-D) Immunofluorescence staining of tubulin and GFP in parasites overexpressing N-terminal deletion and non-Ca²⁺-binding mutant of PfCen1-GFP from pFIO+. Percentage of cells containing ECCAs (arrows) indicated. All images are MIP. DNA stained with Hoechst. Scale bars; 1 μm. (E) Our findings suggest that centrin accumulation at the centriolar plaque from a pre-mitotic diffuse cytoplasmic pool depends on LLPS, which might be promoted by cellular signaling events like calcium levels and a nucleating factor at the centrosome. Parasite is shown without host red blood cell for clarity.