| References: |
ES9-17 is a CME inhibitor a, ES9-17 inhibited recruitment of the plasma membrane-localized BRI1-GFP to the BFA body. Samples were pretreated with 50 µM CHX for 1 h and treated with either DMSO (indicated as 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 Pep-1. The internalization of PEPR1-GFP was followed until 90 min post elicitation. Scale bar, 20 µm. c, Histograms representing the measured endocytic foci lifespan of CLC1-GFP (CLC1:CLC1-GFP/Col-0) in the presence of ES9-17 (30 µM) or DMSO (Ø); n = 1,276 and 1,069 measurements from ten and four seedlings, respectively. Kymographs represent a line trace (horizontal axis), over a time period (vertical axis) taken from a time lapse (2 frames s–1) of Arabidopsis root epidermal cells that illustrates the lifespan of endocytic foci labeled by CHC1-GFP. Scale bar, 10 s. The experiments in a–c were repeated independently three times with similar results. The life time distribution graph was generated by grouping the events into specific categories, annotated as, for example, 0 ≤ x < 10: the category holding those events with a life time (x) being larger or equal (≤) than 0 s and smaller (<) than 10 s. |
