Figure 5.
Tub1 optimizes Dyn1 pathway function.(a) Schematics of GFP- and mRUBY3-tubulin fusion constructs used to visualize microtubules in the indicated single isotype and control cells. (b) Percentage of kar9Δ cells with the indicated tubulin isotypes that display properly aligned anaphase spindles. (c) Percentage of cells displaying Dynein-mediated spindle movements during 10-min observation. (d) Frequency of Dynein-mediated spindle movements in individual cells. (e) Frequency of astral microtubule contact with the bud cell cortex in individual cells. (f) Dynein-mediated spindle movements normalized by microtubule–cortex interactions in individual cells. (g) Spindle translocation per Dynein-mediated event. (h) Duration of Dynein-mediated spindle movements. (i) Astral microtubule length at initiation of Dynein-mediated spindle movements. (j) Spindle translocation per Dynein-mediated event binned by astral microtubule length. (k) Dynein-mediated spindle movements normalized by microtubule–cortex interactions in epothilone B (EpoB)–treated cells. (l) Spindle translocation per Dynein-mediated event binned by astral microtubule length in EpoB-treated cells. For panels b and c, graphs show mean ± SEM from three trials. In panel b, for Tub1 only, n = 86, 92, and 150; for WT-Tub1, n = 83, 107, and 117; for WT-Tub3, n = 98, 76, and 163; for Tub3 only, n = 66, 113, and 158. In panel c, n = 40, 30, and 30 cells for each genotype. For panels d–i, graphs show mean ± SEM from 90–100 cells observed over three trials, where n = 40, 20–30, and 30 for each genotype. For panels g–i, the total number of events recorded is 128, 92, 65, and 36 for Tub1 only, WT-Tub1, WT-Tub3, and Tub3 only, respectively. In panels j–l, graphs show mean ± SEM; panel j includes a total of 33 and 10 Dynein events from three trials for Tub1-only and Tub3-only cells, and panel l includes a total of 56 and 22 Dynein events from two trials for Tub1-only and Tub3-only cells, respectively; for panel k, n = 40, 30, 30, and 40 cells analyzed from two trials for Tub1 only, WT-Tub1, WT-Tub3, and Tub3 only, respectively. *, P ≤ 0.05; **, P ≤ 0.01; and ***, P ≤ 0.001 by unpaired, two-tailed Student’s t test. MT, microtubule.

Tub1 optimizes Dyn1 pathway function. (a) Schematics of GFP- and mRUBY3-tubulin fusion constructs used to visualize microtubules in the indicated single isotype and control cells. (b) Percentage of kar9Δ cells with the indicated tubulin isotypes that display properly aligned anaphase spindles. (c) Percentage of cells displaying Dynein-mediated spindle movements during 10-min observation. (d) Frequency of Dynein-mediated spindle movements in individual cells. (e) Frequency of astral microtubule contact with the bud cell cortex in individual cells. (f) Dynein-mediated spindle movements normalized by microtubule–cortex interactions in individual cells. (g) Spindle translocation per Dynein-mediated event. (h) Duration of Dynein-mediated spindle movements. (i) Astral microtubule length at initiation of Dynein-mediated spindle movements. (j) Spindle translocation per Dynein-mediated event binned by astral microtubule length. (k) Dynein-mediated spindle movements normalized by microtubule–cortex interactions in epothilone B (EpoB)–treated cells. (l) Spindle translocation per Dynein-mediated event binned by astral microtubule length in EpoB-treated cells. For panels b and c, graphs show mean ± SEM from three trials. In panel b, for Tub1 only, n = 86, 92, and 150; for WT-Tub1, n = 83, 107, and 117; for WT-Tub3, n = 98, 76, and 163; for Tub3 only, n = 66, 113, and 158. In panel c, n = 40, 30, and 30 cells for each genotype. For panels d–i, graphs show mean ± SEM from 90–100 cells observed over three trials, where n = 40, 20–30, and 30 for each genotype. For panels g–i, the total number of events recorded is 128, 92, 65, and 36 for Tub1 only, WT-Tub1, WT-Tub3, and Tub3 only, respectively. In panels j–l, graphs show mean ± SEM; panel j includes a total of 33 and 10 Dynein events from three trials for Tub1-only and Tub3-only cells, and panel l includes a total of 56 and 22 Dynein events from two trials for Tub1-only and Tub3-only cells, respectively; for panel k, n = 40, 30, 30, and 40 cells analyzed from two trials for Tub1 only, WT-Tub1, WT-Tub3, and Tub3 only, respectively. *, P ≤ 0.05; **, P ≤ 0.01; and ***, P ≤ 0.001 by unpaired, two-tailed Student’s t test. MT, microtubule.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal