Carbohydrate-Binding Non-Peptidic Pradimicins for the Treatment of Acute Sleeping Sickness in Murine Models
Fig 3
PRM-S binding and fluid-phase endocytosis analysis.
PRM-S binding to parasite surface VSGs was evaluated by competition labelling assays using HHA, a lectin that binds to VSGs and is rapidly endocytosed [5] or CV-N, a lectin that binds to the surface coat of the trypanosomes. Labelling was measured by FACS at 0 min, 10 min and 60 min of incubation at increasing PRM-S concentrations and visualized by 3D microscopy. (A and B) Quantification (A) and images (B) of the labelling with HHA-FITC (1 μg/ml) in the presence of PRM-S. (C and D) Fluid-phase endocytosis analysis using Alexa Fluor 488-labelled dextran 10,000 in the absence (C) or in the presence (D) of HHA (1 μg/ml). (E and F) Quantification (E) and images (F) of the labelling with CV-N-FITC (0.6 μg/ml) in the presence of PRM-S. Bars, 10 μm. The asterisks show significant differences calculated by the Student’s t-test (n = 3). *, p < 0.05, **, p < 0.005 and ***, p < 0.0005 vs the parental strain.