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. 2016 Feb 1;12(2):e1005436. doi: 10.1371/journal.ppat.1005436

Fig 3. Loss of pentamidine inhibition and gain of resistant parasites by a TbAQP Leu264Arg mutation.

Fig 3

(A) Light scattering assay with yeast protoplasts expressing TbAQP2 Leu264Arg in a 300 mM hypertonic glycerol gradient. Data taken in the absence of pentamidine are labeled red, those taken in the presence of 50 μM pentamidine are blue, and protoplasts without an AQP are black. The error bars denote S.E.M. from three independent experiments. (B) Western blot analysis showing inducible expression of GFP-TbAQP2 Leu264Arg (GFPAQP2L264R). (C) Immunofluorescence microscopy analysis of T. brucei expressing GFPAQP2L264R. Scale-bar, 5 μm. (D) Dose–response curve for pentamidine in aqp2-null stains re-expressing wild-type GFP-TbAQP2 or GFPAQP2L264R. EC50 values are indicated. Error bars indicate SD from two independent clones assessed in triplicate assays.