Extended Data Fig. 9: Loss of CGP doesn’t affect parasite replication or secretory organelles.

a, Quantification of the percentage of vacuoles with loss of CGP signal. loxPcgp-Halo parasites were treated with 50 nM rapamycin for 24 h, 48 h, 72 h, and 96 h. At least 100 vacuoles were counted in each 3 biologically independent replicates. Dots represent the mean of each replicate. b, Parasites lacking cgp showed a normal intracellular replication. For each condition, at least 100 vacuoles were counted in each 3 biologically independent replicates. Replication assays were performed after 96 hours incubation with or without 50 nM rapamycin. c, The gliding length (i) and average gliding speed (ii) of parasites capable of gliding (helical or circular movement) were measured by tracking 18 parasites after 72 hours post induction with rapamycin. Motility was analysed by manual tracking the parasites using Icy software. Data presented as Mean + SEM. Dots represent each parasite tracked. d-f) Localisation of apicoplast, microneme and rhoptry proteins is not affected upon deletion of cgp. Parasites were pre-treated ± 50 nM rapamycin for 1 hour and imaged 72 hours later. HSP60: marker for apicoplast. MIC2, MIC6, and MIC8: markers for microneme proteins. ROP1 and ROP2-4: markers for rhoptry proteins. Scale bar, 5 μm. g, Microneme secretion assay performed on wildtype (WT) parasites and loxPcgp-Halo after 72 hours incubation with or without 50 nM rapamycin. Triangles indicate the unprocessed (blue) and processed (red) form of MIC2. This assay was performed in 3 biological replicates. Quantifications are shown in Fig. 3h. Statistics: Unpaired two tailed Student’s t-test was calculated. Colour-coded P values represent the condition compared. Data were presented in graphs as Mean + SD unless otherwise specified.