Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of PHIL1

Protein (function)
PhIL has been identified in Toxoplasma gondii as an inner membrane complex (IMC) protein. PhIL1 is localized mainly with IMC and the apical cap of T. gondii tachyzoites, which is an apical structure linked to the IMC membrane that covers the parasite’s apical protrusion.

Phenotype
Localization of the GFP-tagged PHIL1 protein at the periphery of merozoites, ookinetes and sporozoites.

Additional information
Concavin-GFP expressing sporozoites showed a rapid recovery of the fluorescence signal after a bleached spot was introduced by a high energy laser suggesting high lateral diffusion of concavin-GFP. In contrast, in bleached PHIL1-GFP sporozoites there was no detectable recovery as expected from a protein anchored in the sub-pellicular network. These data suggest that concavin-GFP is not associated to the subpellicular network or another stable cytoskeletal structure.
Next, we performed super-resolution (STED) co-localization experiments with antibodies against concavin-GFP, CSP and PHIL1-GFP. STED imaging showed that the signals of anti-GFP antibodies detecting PHIL1-GFP and antibodies against CSP were spatially separated. In contrast, anti-GFP antibodies detecting concavin-GFP co-localized with anti-CSP antibodies. Similarly, antibodies recognizing CSP but stained with two different colours also co-localized. This suggests that concavin-GFP is localized closer to the plasma membrane than PHIL1, probably within the alveolar space between IMC and plasma membrane or at the inner leaflet of the plasma membrane. 

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