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. Author manuscript; available in PMC: 2022 Feb 3.
Published in final edited form as: Nat Chem Biol. 2019 Apr 22;15(6):641–649. doi: 10.1038/s41589-019-0262-1

Figure 3. ES9-17 is not a protonophore.

Figure 3

(a) Structure of ES9-17 (14). (b) Dose-response of ES9-17-mediated FM4-64 uptake inhibition. The data are plotted as the ratio of the intracellular over the plasma membrane (PM) FM4-64 fluorescence intensities. The EC50 is 13 μM. At least 3 cells were measured per seedling, n=6 seedlings per treatment. (c) Strongly reduced FM4-64 uptake (2 μM, 30 min) in Arabidopsis root epidermal cells in the presence of ES9-17 (30 μM) when compared to the DMSO (Ø) control. The samples were pretreated with either ES9-17 (30 μM) or DMSO for 30 min. (d) Measurements of the ATP concentration in wild type Arabidopsis PSB-D cell cultures treated for 30 min with ES9 (10 μM), ES9-17 (30 μM) and ES9-6 (50 μM), showing ATP concentration relative to the DMSO control. Error bars indicate standard error of the mean of three biological replicates. (e) Confocal images of Arabidopsis epidermal root cells stained with Lyso Tracker Red DND 99 (30 min) and treated additionally for 30 min with DMSO (Ø), ES9 (10 μM) or ES9-17 (30 μM). Scale bars, 10 μm.