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. 2014 Feb 25;9(2):e89771. doi: 10.1371/journal.pone.0089771

Figure 2. PfRab7 localizes to the same compartment as the retromer cargo-selective complex.

Figure 2

(A) Effect of the DD-stabilizing agent trimethroprim (TMP) on the steady-state levels of DD-mCherry-PfRab7 in clonal parasite lines D9 (one expression cassette per genome) and G9 (two expression cassettes per genome). 3D7 is the untransfected parental parasite line. Upper: Anti-mCherry immunoblot. Lower: The membrane was reprobed with anti-glycerophosphoester phosphodiesterase (PfGDPD) antibodies for a loading control. The sizes of protein markers in kDa are indicated at left. (B) Images of DD-mCherry-PfRab7 in live clone G9 parasites. Stages shown are ring (R), trophozoite (T), schizont (Sz), and extracellular merozoite (M). (C) Live parasites expressing DD-mCherry-PfRab7ΔCC and DD-mCherry. (D) Live parasites co-expressing PfVps35-YFP and DD-mCherry-PfRab7. (E) Live parasites co-expressing PfVps35-YFP and DD-mCherry-PfRab6. (F) Live parasites expressing DD-mCherry-PfRab7 mutants. T22N and N125I are predicted to be “GDP-locked” and Q67L is predicted to be “GTP-locked”. 5′ UTR sequences driving expression are pfrab7 for N125I and Q67L and pfapp (aminopeptidase P) for T22N. In panels B, C and F, Hoechst 33342 fluorescence (DNA) is pseudocolored green and in panels D and E YFP fluorescence is pseudocolored green. In panels B–F, parasites were cultured in the presence of 10 µM trimethoprim. mCherry (mC) fluorescence is pseudocolored red. Scale bars, 2 µm.