Fig. 5
P. falciparum RASP2 is necessary for invasion. a Strategy for conditional excision of the pfrasp2 gene using the DiCre system. b PCRs showing efficient excision of the pfrasp2 locus upon addition of rapamycin with primers 3181/2911 (non-excised), 3181/2912 (excised). c IFA on iKO-PfRASP2-HA3 schizonts ± rapamycin. d Immunoblot on schizont lysates from the p230p DiCre (parental line) and iKO-PfRASP2 mutant line ± rapamycin. PfAldolase, loading control. e Left: Growth curves (parasitaemias) of p230p DiCre (Ctrl) and iKO-PfRASP2 mutant ± rapamycin shows that PfRASP2-depleted parasites are unable to proliferate. Right: Giemsa stainings illustrating the development and reinvasion of iKO-PfRASP2 merozoites (48 h) in the absence of rapamycin and their accumulation at the surface of RBCs in the presence of rapamycin treatment. f Graphical summary of egress data of control (-rapa) and rapamycin-treated iKO-PfRASP2 schizonts following removal of C2. Data collected from 8 control movies (-rapa) and 11 movies in the presence of rapamycin. Number of egress events normalized as a percentage of control (-rapa) considered as 100% egress here. g Invasion rate of Ctrl and iKO-PfRASP2 parasites treated ± rapamycin. Synchronized cultures were treated for 6 h at ring stage with rapamycin or DMSO and were quantified by flow cytometry (100,000 RBCs were analysed for each time point) over 2 cycles (see methods). (e, g) Values represent means ± SD (n = 3) from a representative experiment out of two independent assays. Source data are provided as a Source Data file
