Fig. 7.

CPH1 mediates important interaction with hypothetical unknown conoid proteins. a Analyses of interactions with CIP1, CIP2, and CIP3. Left panel: domain analysis as determined by InterPro, and domain names as shown in Fig. 5c. CRISPR fitness scores were derived from ref.³². Right panels: intracellular and extracellular parasites were treated, fixed, and stained as described in Fig. 6b, c. Scale = 2 μm. b PLA for interaction of CPH1 with the CIPs was performed as described in Fig. 6d. Scale = 2 μm. a, b Representative of three or more experiments with similar outcomes. c CIP1, CIP2, and CIP3 were sequentially deleted, generating double and triple knockouts. Plaque sizes and numbers are plotted (N > 50 plaques). Mean ± SEM for 3 experiments with 3 replicates for each (n = 9). ***P < 0.0001, Kruskal–Wallis test with Dunn’s correction for multiple comparison. For details on generation of the parasite lines, see Supplementary Fig. 7. d Parasites were treated with 3 µM A23187 to protrude the conoid, prior to EM processing. Four major categories of conoid structure were identified with examples shown on left: (1) normal conoid from TIR1 parental line; (2) loss of the pre-conoid ring from ∆cip1∆cip2; (3) disrupted conoid, and (4) collapsed conoid from ∆cip1∆cip2∆cip3 (colored for the category numbers). Ratios of different categories were calculated from two independent experiments, and averages were shown (N = 20 conoids for each group). Scale = 500 nm. ns not significant