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. 2006 Aug 28;174(5):665–675. doi: 10.1083/jcb.200601062

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Copyright © 2006, The Rockefeller University Press

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Figure 6. — mps2-381 mutants are defective in SPB duplication. (A) Wild-type (SLJ001) and mps2-381 (SLJ2006) were grown to midlog phase at 23°C and then shifted to 37°C for 4 h. Flow cytometric analysis of DNA content showed that although wild-type cells are haploids (1N and 2N) at 23°C, the mps2-381 mutants are diploids (2N and 4N) and arrest at 37°C in mitosis. The percentage of large-budded cells was also determined (n = 200). (B) Indirect immunofluorescence microscopy revealed that mps2-381 cells arrested at 37°C with monopolar spindles: a single microtubule array (anti-Tub1; green) nucleated from one SPB (anti-Tub4; red) associated with one DNA mass (DAPI; blue). Bar, 5 μm. (C) Nuclei from large-budded cells were examined by EM to further analyze SPB number and mitotic spindle structure in mps2-381 cells arrested at 37°C. Thin section images of two representative SPBs are shown; both SPBs are on deep nuclear invaginations and lack any recognizable half-bridge structure. Bar, 0.2 μm.