Fig. 2: Efficient Rapamycin-induced excision of floxed NBPXa. | Nature Communications

Fig. 2: Efficient Rapamycin-induced excision of floxed NBPXa.

From: Sequential roles for red blood cell binding proteins enable phased commitment to invasion for malaria parasites

Fig. 2

a Schematic depicting excision of 8.5kbp fragment between LoxP sites (L) upon Rapamycin (Rap) treatment of NBPXa cKO parasites. Diagnostic primer positions noted in blue (P1, P2, P3). b Diagnostic PCRs showing outcome of DMSO (D) or Rap (R) treatment of NBPXa cKO parasites. Primers P1/P2 identified non-excised parasites, P2/P3 detected successful excision and control bands amplify an unrelated locus. c Western blot showing loss of ~325 kDa HA tagged NBPXa in Rap treated parasites. Anti-PfHSP70 used as a loading control. d IFAs showing ablation of NBPXa expression for Rap treated parasites. Parasites labelled with a rat anti-HA and rabbit anti-MSP119 as a marker for mature, segmented schizonts. Scale bars = 5 μm. e Growth of NBPXa cKO parasites in human (left) or macaque (right) RBCs measured at 24 and 48 h after treatment with Rap or DMSO. Data shown are the mean of 5 independent experiments for human and 2 experiments for macaque cells. Error bars +/- SEM. Source data for (b) and (c) and (e) are provided as a Source Data file.

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