Figure S5.
Exogenous copy of TUB1 and co-overexpression of α- and β-tubulin are tolerated. (A) Schematic of the CEN-based additional copy of TUB1.(B) Normalized doubling times of wild-type or tub1Δ/TUB1 with an empty vector or additional copy of TUB1 on a CEN plasmid. For each genotype, four technical replicates of two biological replicates were used across three independent experiments. Circles represent cultures in separate wells and triangles are the means of each day. Colors indicate independent experiments. Asterisk indicates P < 0.05 between wild type and mutant based on t test after one-way ANOVA. Bars are mean ±95% CI. (C) Tenfold dilution series of indicated strains were spotted onto rich medium or rich medium supplemented with benomyl. Cells were grown at the indicated temperature for the indicated number of days. (D) Histogram of all astral microtubule lengths from time-lapse imaging of wild-type cells with an empty vector or an extra copy of TUB1. Data are from at least three separate experiments for each genotype, and a total of at least 26 cells were analyzed for each genotype. (E) Polymerization rates of astral microtubules. Each dot represents a single polymerization event, and dots are colored by experimental day. Bars are median ± 95% CI. (F) Paired molecules of α- or β-tubulin per cell for each genotype. Protein mass (ng) was converted to molecules per cell. Data represent four independent experiments with two biological replicates, and each dot is mean molecules per cell of the four technical replicates. Dots are colored by experiment. (G) Table of astral microtubule dynamics and soluble tubulin levels in diploid cells + TUB1 or + ev. Values in bold have P < 0.05 from wild type + ev, based on a t test. a, Median values (95% CI); b, Mean values ±95% CI; c, Mean values ±95% CI. (H) Representative Western blot of β- and α-tubulin during TUB1 and TUB2 co-induction. Blots were probed for α- or β-tubulin and Zwf1 (G6PD) as a loading control. (I) Quantification of TUB1 and TUB2 co-induction for both α- and β-tubulin across overexpression. Dots represent mean ± SD from three independent induction experiments. (J) Quantification of cells forming colonies after TUB1 and TUB2 induction or with empty vector. Dots represent mean ± SD from three independent induction experiments. Source data are available for this figure: SourceData FS5.

Exogenous copy of TUB1 and co-overexpression of α- and β-tubulin are tolerated. (A) Schematic of the CEN-based additional copy of TUB1.(B) Normalized doubling times of wild-type or tub1Δ/TUB1 with an empty vector or additional copy of TUB1 on a CEN plasmid. For each genotype, four technical replicates of two biological replicates were used across three independent experiments. Circles represent cultures in separate wells and triangles are the means of each day. Colors indicate independent experiments. Asterisk indicates P < 0.05 between wild type and mutant based on t test after one-way ANOVA. Bars are mean ±95% CI. (C) Tenfold dilution series of indicated strains were spotted onto rich medium or rich medium supplemented with benomyl. Cells were grown at the indicated temperature for the indicated number of days. (D) Histogram of all astral microtubule lengths from time-lapse imaging of wild-type cells with an empty vector or an extra copy of TUB1. Data are from at least three separate experiments for each genotype, and a total of at least 26 cells were analyzed for each genotype. (E) Polymerization rates of astral microtubules. Each dot represents a single polymerization event, and dots are colored by experimental day. Bars are median ± 95% CI. (F) Paired molecules of α- or β-tubulin per cell for each genotype. Protein mass (ng) was converted to molecules per cell. Data represent four independent experiments with two biological replicates, and each dot is mean molecules per cell of the four technical replicates. Dots are colored by experiment. (G) Table of astral microtubule dynamics and soluble tubulin levels in diploid cells + TUB1 or + ev. Values in bold have P < 0.05 from wild type + ev, based on a t test. a, Median values (95% CI); b, Mean values ±95% CI; c, Mean values ±95% CI. (H) Representative Western blot of β- and α-tubulin during TUB1 and TUB2 co-induction. Blots were probed for α- or β-tubulin and Zwf1 (G6PD) as a loading control. (I) Quantification of TUB1 and TUB2 co-induction for both α- and β-tubulin across overexpression. Dots represent mean ± SD from three independent induction experiments. (J) Quantification of cells forming colonies after TUB1 and TUB2 induction or with empty vector. Dots represent mean ± SD from three independent induction experiments. Source data are available for this figure: SourceData FS5.

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