Fig. 3.

mTOR inhibition by other means does not phenocopy the antiplasmodial effect of torins. (A and B) Effect of PP242 on inhibition of liver stage infection. Infected HepG2 cells were treated with 2.5 uM PP242 2 h after spozoroite addition, and the infection was analyzed by flow cytometry 48 h later, as in Fig. 1. Data represent mean ± SD three independent experiments. (C) Effect of PP242 on blood stage infection. Synchronized P. falciparum 3D7 rings were treated with 2.5 μM PP242 or DMSO (identical control data set as in Fig. 2A), and reinvasion was analyzed after SybrGreen labeling of parasite DNA 48 h later. Data represent mean ± SD of three independent experiments. (D–F) Effect of siRNA-mediated knockdown of raptor or mTOR on liver stage infection. Validated siRNA pools were reverse-transfected into HepG2 cells, which were infected with sporozoites 2 d later and analyzed by flow cytometry as described above or with immunofluorescence analysis (IFA) 24 h after sporozoite addition. (F) Representative images with PbHSP7 (green), UIS4 (red), and DAPI (blue).