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. 2019 Feb 26;8:e42182. doi: 10.7554/eLife.42182

Figure 1. In S. pombe spindle length scales with cell size but total spindle duration is kept constant.

(A) Brightfield images of S. pombe wild-type cells (central panel) and cell cycle mutants wee1-50 (left panel) and cdc25-22 (right panel). Cells reach their maximum cell length at mitosis/cell division, represented by the septum (asterisk). Scale bar, 5 µm. (B) Time-lapse images of wee1-50, wild-type (wt) and cdc25-22 cells expressing mCherry-Atb2 (tubulin) and Mis12-GFP from spindle assembly to spindle breakdown (asterisk). Dotted line corresponds to transition from phase I to II (prophase to metaphase). Dashed line corresponds to transition from phase II to III (metaphase to anaphase). Arrowheads indicate the last time point of phase II or III, corresponding to spindle length plotted in (D) and Figure 1—figure supplement 1. Each frame corresponds to 1 min interval. Scale bar, 5 µm. (C) Comparative plot of spindle length dynamics of wee1-50 (red curves, n = 20), wild-type (green curves, n = 20) and cdc25-22 cells (blue curves, n = 20). Bold curves correspond to mean spindle dynamics. As in (B) dotted and dashed lines display phase transitions. (D) Final spindle length plotted against cell length (wee1-50: n = 53, wt: n = 61, cdc25-22: n = 60). (E) Total spindle duration plotted against cell length (wee1-50: n = 53, wt: n = 61, cdc25-22: n = 60). Data in (D–E) was fitted by linear regression (dashed lines), showing the regression coefficient R2 and the slope m. (F) Boxplot comparison of anaphase B spindle elongation velocities (v) in wee1-50, wild-type and cdc25-22 cells. Data from n cells was collected from three independent experiments. P values were calculated by Mann-Whitney U test.

Figure 1—source data 1. Mean values of spindle length and dynamics.
Mean values (bold) and corresponding standard deviations of cell length, total spindle duration and spindle length, duration and velocity for each mitotic phase: phase I (prophase), phase II (metaphase-anaphase A) and phase III (anaphase B) of wee1-50 (n = 53), wild-type (n = 61) and cdc25-22 cells (n = 60). Data obtained from n analyzed cells was collected from three independent experiments.
elife-42182-fig1-data1.docx (13.3KB, docx)
DOI: 10.7554/eLife.42182.005

Figure 1.

Figure 1—figure supplement 1. Metaphase spindle length plotted against maximum cell length.

Figure 1—figure supplement 1.

Data was fitted by linear regression (dashed line), showing the regression coefficient R2 and the slope m. Data obtained from n analyzed cells (wee1-50: n = 53, wt: n = 61, cdc25-22: n = 60) was collected from three independent experiments.
Figure 1—figure supplement 2. Boxplot comparison of spindle elongation velocity in prophase (I), metaphase (II) and anaphase B (III).

Figure 1—figure supplement 2.

P values were calculated by Mann-Whitney U test. Data obtained from n analyzed cells was collected from three independent experiments.