Figure 2: ES9 inhibits CME dynamics and organelle movement in the cytoplasm.
(a) Kymographs representing a line trace (horizontal axis) over a time period (vertical axis, 5 min), taken from the respective spinning disc movies (2 f s−1) of Arabidopsis root cells and illustrating the life times of endocytic spots labelled by TML-GFP for the control treatment (DMSO, Ø), 10 μM ES9, 50 μM tyrphostinA23 (TyrA23), and 50 μM TyrA51. (b) Boxplot representation of measured endocytic spot life times. n=1646, 171, 121, and 868 measurements. (c) Boxplot representation of kymograph-based life time of foci at the plasma membrane in the presence of 10 μM ES9 for green fluorescent protein (GFP)-tagged TPLATE, clathrin light chain (CLC), clathrin heavy chain (CHC), the adaptor protein complex-2 (AP-2), represented by the AP2A1, AP2M, and AP2S subunits, and the dynamin DRP1A. n=204, 125, 142, 243, 60, 150, 176, 121, and 247 measurements. (d) Visualization of the Golgi (ST-mRFP) and the trans-Golgi network (TGN, VHA-a1-mRFP) in Arabidopsis root cells treated with 10 μM ES9, 50 μM TyrA23, 50 μM TyrA51, and mock (DMSO, Ø) (5 min). Images are composed of six differentially coloured images taken with a 10-s interval. Movement is illustrated by the presence of the different colours, whereas white indicates static compartments. Boxplot centre lines show the medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles; outliers are represented by dots. Scale bar, 5 μm.
