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. 2022 Dec 19;25(1):56–67. doi: 10.1038/s41556-022-01035-2

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a, Kar9-mNeonGreen fluorescence intensity levels in Kar9 self-association interface mutants and bik1Δ at 25 °C at a single astral microtubule plus-end (normalized median with 95% confidence intervals). P values determined by Welch’s t-test. For cumulative fluorescence intensity on all plus-ends, see Extended Data Fig. 7. b, Kar9 asymmetry index (Methods) quantifying Kar9-mNeonGreen distribution between astral microtubule tips on both sides of metaphase spindles at 25 °C (median with 95% confidence interval). P values determined by Welch’s t-test. Example for a weakly symmetric kar9-αβγ⁻ bik1Δ cell with labels for quantified intensities is shown on the right. Scale bar, 3 μm. Blue values in a and b indicate n = cells analysed/number of biological replicates. c, Micrographs showing Kar9-mNeonGreen tracking the plus-end of a shrinking astral microtubule in wild type (WT, arrowheads mark the period of shrinkage) and failure to do so in a kar9-αβγ⁻ bik1Δ cell at 25 °C (the asterisk indicating loss of Kar9 punctum). Scale bars, 3 μm and 1 μm (in zooms). See also Supplementary Movies 7 and 8. d, Astral microtubule length profiles for wild-type, kar9-αβγ⁻, bik1Δ and kar9-αβγ⁻ bik1Δ cells (moving average over three timeframes of distance between Kar9-mNeonGreen and Spc42-mCherry). Profiles persisting for the whole length of the movie are displayed in grey; profiles that end owing to loss of punctum (indicated with arrowhead) in red. e, Frequency of cells losing Kar9-mNeonGreen punctum on astral microtubule plus-ends over 175 s (error bars are Wilson/Brown 95% confidence intervals determined from binomial distribution). P values are determined by two-proportion z-test. Blue values indicate n (cells analysed, single experiment). f,g, Estimated rates of Kar9-mNeonGreen fluorescence change (f) and astral microtubule length change (g) for growing (blue) and shrinking (red) microtubules. Symbols indicate estimates and lines their 95% confidence intervals. Colour of P values (two-sided t-test) and n (number of timeframes considered for the model, single experiment) matches the growth/shrinkage category they refer to. For details, see Methods and Extended Data Fig. 8. Source numerical data available in source data.

Source data