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. 2008 Sep 15;52(11):4030–4036. doi: 10.1128/AAC.00964-08

FIG. 3.

FIG. 3.

Correlation between sitamaquine uptake and the pH of acidic organelles in different Leishmania strains. (A) [14C]sitamaquine uptake in different Leishmania strains. Accumulation assays were determined as described in the legend to Fig. 2. Parasites were pretreated with 20 mM NH4Cl for 1 min and with the ionophores nigericin and monensin at 10 μM for 10 min at 28°C. [14C]sitamaquine uptake was determined for 5 min as described in Materials and Methods. (B) Determination of the sensitivity to sitamaquine in L. tropica and L. donovani strains by Alamar Blue assay. Statistical significance using Student's t test was considered for P values of <0.05. Values for L. donovani L82 versus those of L. donovani dd8, L. tropica LRC, and L. tropica k27 were significantly different (P < 0.05, P < 0.0005, and P < 0.0004, respectively). Values for L. donovani dd8 versus those of L. tropica LRC and L. tropica k27 were significantly different (P < 0.001 and P < 0.02, respectively). (C) Effect of NH4Cl on Lysotracker Green fluorescence by flow cytometry. A representative graph is shown. Fluorescence of L. donovani L82 after incubation with 100 nM Lysotracker Green for 10 min is shown in black. Decrease in fluorescence produced by 20 mM NH4Cl at 1, 5, and 8 min is represented by the gray lines (a, b and c, respectively). The dotted line corresponds to parasites’ autofluorescence. Data are the means ± standard deviations of three independent experiments. SQ, sitamaquine.