Figure 4: Increased metaphase spindle length reduces anaphase transition time in cut7Δpkl1Δ cells. | Nature Communications

Figure 4: Increased metaphase spindle length reduces anaphase transition time in cut7Δpkl1Δ cells.

From: Kinesin-5-independent mitotic spindle assembly requires the antiparallel microtubule crosslinker Ase1 in fission yeast

Figure 4

(a) Time-lapse images of cut7Δpkl1Δ and cut7Δpkl1Δase1OE cells expressing mCherry-Atb2 and Sid4-GFP from mitosis onset to spindle breakdown. Each frame corresponds to 3 min interval. (b) Comparative plots of spindle length dynamics of cut7Δpkl1Δ (red curves; n=33) and cut7Δpkl1Δase1OEcells (orange curves; n=15). Dotted vertical line corresponds to phase I–II transition; dashed vertical line corresponds to phase II–III transition. (c) Time-lapse images of wild-type, cut7Δpkl1Δ and cut7Δpkl1Δase1OE cells expressing mCherry-Atb2 and Cdc13-GFP from mitosis onset to beginning of anaphase. Each frame corresponds to 3 min interval. (d) Bar plot showing metaphase spindle length at anaphase transition, reported by Cdc13-GFP degradation, of wild-type (green; n=33), ase1OE (light blue; n=28), cut7Δpkl1Δ (red; n=25) and cut7Δpkl1Δase1OE (orange; n=30). *P<0.001. (e) Bar plot showing anaphase transition time of wild-type (green; n=33), ase1OE (light blue; n=28), cut7Δpkl1Δ (red; n=25) and cut7Δpkl1Δase1OE (orange; n=30). *P<0.01. (f) Time-lapse images of cut7Δpkl1Δ and cut7Δpkl1Δcls1OEcells expressing mCherry-Atb2 and Sid4-GFP from mitosis onset to spindle breakdown. Each frame corresponds to 3 min interval. (g) Comparative plot of spindle length dynamics of cut7Δpkl1Δ (red curves; n=31) andcut7Δpkl1Δcls1OEcells (orange curves; n=19). Dotted vertical line corresponds to phase I–II transition; dashed vertical line corresponds to phase II–III transition. (h) Time-lapse images of wild-type, cut7Δpkl1Δ and cut7Δpkl1Δcls1OE cells expressing mCherry-Atb2 and Cdc13-GFP from mitosis onset to beginning of anaphase. Each frame corresponds to 3 min interval. (i) Bar plot showing metaphase spindle length at anaphase transition, reported by Cdc13-GFP degradation, of wild-type (green; n=37), cls1OE (light blue; n=23), cut7Δpkl1Δ (red; n=88) and cut7Δpkl1Δcls1OE (orange; n=97). *P<0.001. (j) Bar plot showing the time from MT nucleation to Cdc13-GFP degradation of wild-type (green; n=37), cls1OE (light blue; n=23), cut7Δpkl1Δ (red; n=88) and cut7Δpkl1Δcls1OE cells (orange; n=97). *P<0.05. (k) Model for spindle assembly and transition to anaphase (see text for details). Data show mean±s.d. and Student’s t-test P value. Scale bars, 2 μm.

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