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. 2018 Apr 19;1(2):e201800054. doi: 10.26508/lsa.201800054

Figure 6. β-CTT function requires negatively charged amino acids.

Figure 6.

(A) Amino acid sequences of β-CTTs of mutant yeast strains characterized in Fig 6. The native (TUB2) allele provided for comparison. (B) Representative images of preanaphase yeast strains expressing microtubules labeled with GFP-Tub1 treated with either 1.5 μM nocodazole or DMSO as a control. Arrowheads indicate the plus ends of astral microtubules (aMTs). (C) Percent of cells exhibiting aMTs after exposure to 1.5 μM nocodazole or DMSO for 1 h. Bars represent the pooled mean ± standard error of the proportion of four separate experiments pooled. Each group is composed of n ≥ 735 cells. **P « 0.001. Significance determined by Fisher's exact test. (D) Representative aMT life plots of wild-type and tub2-polyQ cells expressing GFP-labeled tubulin. Microtubule lengths were measured at 3- to 4-s time intervals. (E) Distributions of aMT lengths in living preanaphase cells. Images were collected at 3- to 4-s intervals for 10 min. Results represent pooled data from at least four separate experiments and at least 40 aMTs sampling n ≥ 3,593 time points for each strain. Red bars indicate median values. **P « 0.001. Significance determined by the Mann–Whitney U test. (F) Distributions of aMT polymerization rates, from cells analyzed in (E). (G) Distributions of aMT depolymerization rates, from cells analyzed in (E). (H) Distributions of aMT catastrophe frequency, from cells analyzed in (E).