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. 2023 Aug 17;34(10):ar97. doi: 10.1091/mbc.E23-02-0063

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© 2023 Sherwin et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 4: — Loss of Fin1 impairs Slk19 spindle midzone localization and causes broken spindles in telophase. (A) Slk19 spindle midzone localization is reduced in fin1Δ cells. WT (4207-1-3) and fin1Δ (4207-4-1) cells containing Slk19-GFP and Tub1-mCherry were cultured overnight at 25°C in YPD medium. Asynchronous cells were collected for fluorescence microscopy. The percentage of cells with elongated spindles harboring Slk19 midzone localization was quantified. White dotted boundary represents cell lacking Slk19 midzone localization. Scale bar, 5 μm. The pictures are representative of three experimental repeats (N = 3) where 100 cells were counted for each experiment. The bars represent mean values ± SD. p values were obtained by using a two-tailed unpaired t test, with asterisks indicating ***p < 0.001. (B) Ase1 spindle midzone localization does not change in fin1Δ mutant cells. WT (2567-2-1) and fin1Δ (4205-5-1) cells containing Ase1-GFP and Tub1-mCherry were subjected to the same experiment as in panel A. White dotted boundary represents Ase1 midzone localization in cells with elongated spindles. Scale bar, 5 μm. The pictures and graph represent the quantification of counting 100 cells with elongated spindles for the WT and mutant, respectively. (C) Loss of Slk19 results in broken spindles in cells arrested at telophase. cdc15-2 (870-1-2) and cdc15-2 slk19Δ (870-2-2) cells containing Tub1-GFP were grown to log phase in YPD at 25°C then shifted to 37°C for 3 h to inactivate Cdc15. The percentage of cells with broken spindles was quantified. The yellow arrow points to a broken spindle in a representative cell outlined with white dotted line. Scale bar, 5 μm. The pictures are representative of three experimental repeats (N = 3) where 100 cells were counted for each experiment. The bars represent mean values ± SD. p values were obtained by using a two-tailed unpaired t test, with asterisks indicating **** p < 0.0001. (D) Loss of Fin1 results in broken spindles in cells arrested at telophase. cdc15-2 (4370-4-3) and cdc15-2 fin1Δ (4370-1-2) cells containing Tub1-GFP were subjected to the same experiment as in panel C. Yellow arrows point to broken spindles in representative cells outlined with white dotted lines. Scale bar, 5 μm. The pictures are representative of three experimental repeats. The same method was used for statistical analysis as described in panel C.