Fig. 1.

PfBLEB localizes to the basal complex in asexual-stage parasites. (A) Immunofluorescence of PfBLEB-smV5^Tet shows close proximity of PfBLEB and basal complex marker, PfMORN1. (B) PfBLEB has distinct localization from the inner membrane complex-associated marker, PfGAP45. SR-SIM images shown are single z slices for individual channels and maximum intensity projections of merged channels. (Scale bars, 1 µm.) (C) Removal of ATc results in 87 ± 14% knockdown of PfBLEB-smV5 protein levels. PfBLEB-smV5^Tet parasites were grown in the presence or absence of ATc for 2 cycles, then immunoblot of smV5 tag in late-stage schizonts was used to quantify the level of protein knockdown. Quantification performed via volumetric measurement of fluorescence intensity with the LiCor Odyssey CLx system and fluorescence compared to histone H3 loading control. Means ± SEMs. Unpaired, two-tailed t test. ***P < 0.001. n = 3 technical replicates per experiment, representative of 3 independent experiments. Representative western blot shown in SI Appendix, Fig. S5. (D) Knockdown of PfBLEB via 8xTet/TetR-DOZI system does not affect asexual parasite replication. PfBLEB-smV5^Tet parasites were grown in the presence or absence of ATc for 2 cycles. Parasitemia was measured via flow cytometry of SYBR Green I. Means ± SDs. Error bars smaller than size of marker. n = 3.