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. 2013 Apr 1;24(7):914–922. doi: 10.1091/mbc.E12-11-0812

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© 2013 King et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 1: — Degradation of Swe1p is not affected by osmotic shock in the absence of feedback via Cdc28p. (A) Swe1p and septin-Swe1p levels rise and fall in parallel as cells traverse the cell cycle. Cells containing CDC28^E12K to prevent Swe1p-mediated Cdc28p inhibition (DLY12321) were arrested in G1 with pheromone and released into fresh media at 30°C to traverse the cell cycle. Pheromone was reintroduced 60 min after release to rearrest cells at the next G1. Samples were taken at 15-min intervals and the levels of myc-tagged Swe1p and septin-Swe1p were analyzed by Western blotting with anti-myc (Cdc11p: loading control), and quantitated (graph). (B) Synchrony parameters (budding and nuclear division) for the experiment in (A) were scored for >200 cells per sample. (C) Swe1p and septin-Swe1p levels are unaffected by osmotic shock. Cells were treated as in (A), except that 0.4 M NaCl was added at 45 min. (D) Synchrony parameters for the experiment in (C) show that osmotic stress transiently perturbed bud formation and nuclear division. (E) A single-cycle synchrony experiment as in (A) was performed with strain DLY12034, which does not express septin-Swe1p. (F) Synchrony parameters for (E). (G) Cells were treated as in (E), except that 0.4 M NaCl was added at 45 min. (H) Synchrony parameters for (G).