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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 4. ES9-17 is a CME inhibitor.

Figure 4

(a) ES9-17 inhibited the recruitment of the plasma membrane-localized BRI1-GFP to the Brefeldin A (BFA) body. Samples were pretreated with 50 μM cycloheximide (CHX) for 1 h and treated with either DMSO (Ø) or ES9-17 (30 μM) for 30 min, followed by a combined application of FM4-64 (2 μM) and BFA (50 μM) for an additional 30 min in the presence of the inhibitor. Scale bar, 10 μm. (b) Confocal images of Arabidopsis root epidermal cells expressing PEPR1-GFP. Seedlings were treated with ES9-17 (30 μM) or DMSO (Ø) for 30 min, followed by elicitation with 100 nM Pep1. The internalization of PEPR1-GFP was followed until 90 min post elicitation. Scale bar, 20 μm. (c) Histograms representing the measured endocytic foci life time of clathrin light chain1 (CLC1)-GFP (CLC1::CLC1-GFP/Col-0) in the presence of ES9-17 (30 μM) or DMSO (Ø), n=1276 and 1069 measurements, from 10 and 4 seedlings respectively. Kymographs represent a line trace (horizontal axis) over a time period (vertical axis), taken from a time lapse (2 f/sec) of Arabidopsis root epidermal cells, that illustrate the life times of endocytic foci labeled by CHC1-GFP. Scale bar, 10 sec.