Figure 6.
Cargo capture and dynein activation are coupled. (a) Image from the first frame of a time-lapse video (left) of dynein (green) and dextran (magenta), and the corresponding kymograph (right). The white arrowheads point to dynein and dextran vesicles moving together toward the minus end and are shown separately in the images on the bottom. See also Fig. S5 f for an example of another such event in the same cell. (b) Plot of position versus time of the dextran vesicle (magenta) indicated in a, alongside the intensity of dynein on that vesicle (green), showing a short minus end–directed run of the vesicle upon dynein binding. n = ∼15 cells across two independent repeats. (c) Image from the first frame of a time-lapse video (left) of dynein (green) and EGF (magenta), and the corresponding kymograph (right). The white arrowheads point to dynein and EGF vesicle moving together toward the minus end and are shown separately in the images on the bottom. (d) Plot of position versus time of the EGF vesicle (magenta) indicated in c, alongside the intensity of dynein on that vesicle (green), showing a short minus end–directed run of the vesicle upon dynein-binding. n = ∼15 cells across two independent repeats. (e) Images from time-lapse video of EGF entry and subsequent movement within cells. Time is indicated above the images in min:s. (f) Kymograph of the region indicated by the magenta rectangle in e, with the time-lapse images sampled every 5 s showing entry of EGF and the net movement of endosomes toward the nucleus. (g) Kymograph of the trajectory marked by the white dashed rectangle in f, where the time-lapse images were sampled every 0.5 s. (h) Quantification of the net distance moved by dextran (“Dex”) and EGF endosomes in 1 min. Asterisks indicate significant difference, P = 4 × 10−5, one-way ANOVA, Tukey Kramer post-hoc test. n = 24 dextran endosomes from 12 cells across two independent repeats and 32 EGF endosomes from nine cells across two independent repeats. Error bars represent SD. (i) Schematic of dynein’s cargo search mechanism: stochastic binding of dynein to the MT at a location proximal to the cargo–adaptor complex leads to a short minus end–directed run, which terminates upon the unbinding of dynein. In a, c, and e, “N” marks the location/direction of the nucleus.

Cargo capture and dynein activation are coupled. (a) Image from the first frame of a time-lapse video (left) of dynein (green) and dextran (magenta), and the corresponding kymograph (right). The white arrowheads point to dynein and dextran vesicles moving together toward the minus end and are shown separately in the images on the bottom. See also Fig. S5 f for an example of another such event in the same cell. (b) Plot of position versus time of the dextran vesicle (magenta) indicated in a, alongside the intensity of dynein on that vesicle (green), showing a short minus end–directed run of the vesicle upon dynein binding. n = ∼15 cells across two independent repeats. (c) Image from the first frame of a time-lapse video (left) of dynein (green) and EGF (magenta), and the corresponding kymograph (right). The white arrowheads point to dynein and EGF vesicle moving together toward the minus end and are shown separately in the images on the bottom. (d) Plot of position versus time of the EGF vesicle (magenta) indicated in c, alongside the intensity of dynein on that vesicle (green), showing a short minus end–directed run of the vesicle upon dynein-binding. n = ∼15 cells across two independent repeats. (e) Images from time-lapse video of EGF entry and subsequent movement within cells. Time is indicated above the images in min:s. (f) Kymograph of the region indicated by the magenta rectangle in e, with the time-lapse images sampled every 5 s showing entry of EGF and the net movement of endosomes toward the nucleus. (g) Kymograph of the trajectory marked by the white dashed rectangle in f, where the time-lapse images were sampled every 0.5 s. (h) Quantification of the net distance moved by dextran (“Dex”) and EGF endosomes in 1 min. Asterisks indicate significant difference, P = 4 × 10−5, one-way ANOVA, Tukey Kramer post-hoc test. n = 24 dextran endosomes from 12 cells across two independent repeats and 32 EGF endosomes from nine cells across two independent repeats. Error bars represent SD. (i) Schematic of dynein’s cargo search mechanism: stochastic binding of dynein to the MT at a location proximal to the cargo–adaptor complex leads to a short minus end–directed run, which terminates upon the unbinding of dynein. In a, c, and e, “N” marks the location/direction of the nucleus.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal