Fig. 6
RASP2 plays an essential role in rhoptry secretion. a Immunoblot showing microneme secretion (arrow = processed/secreted TgAMA1) in TATi_TgRASP2-HA3 (Ctrl) and KD-TgRASP2 parasites. Propranolol induced secretion (Sup induced). TgGRA3, loading control. b Immunoblot of iKO-PfRASP2-HA3 culture supernatant (secreted proteins) showing microneme secretion (PfAMA1 p44) in control and PfRASP2-depleted parasites. Late schizonts were allowed to egress for 45 min in RPMI. Free merozoites and culture supernatant were separated by centrifugation and processed for Western blot and probed with rabbit anti-PfAMA1. c IFA on E64 or DMSO-treated iKO-PfRASP2-HA3 parasites ± rapamycin. PfRASP2 depletion does not affect PfAMA1 relocalisation at the surface of merozoites (green arrows). d Left: IFA showing a representative example of an evacuole/rhoptry secretion event in T. gondii (ROP1 staining), Right: Quantification of evacuoles in TATi_TgRASP2-HA3 (Ctrl) and KD-TgRASP2 parasites (±ATc) (One representative experiment out of three independent assays). e STAT6-P assay on the same lines and the complemented cKD-RASP2 line. f Left: IFA showing two representative examples of a rhoptry secretion event in P. falciparum (RAP2 staining). Right: Quantification of secretion events in iKO-PfRASP2 parasites (±rapamycin). Means ± SD for three independent experiments where >2,500 RBCs were analysed. g Ultrastructure of KD-TgRASP2 + ATc tachyzoites shows no defect in rhoptry positioning. m: micronemes; c: conoid. Source data are provided as a Source Data file
