Ndd1 turnover requires Cdk1, Hog1, and three of the GSK3 family kinases.

<p>A) Cells expressing wildtype Ndd1-Flag (<i>GAL1</i>p-Ndd1^wt-Flag) or an allele with 16 S/T-P consensus sites mutated (<i>GAL1</i>p-Ndd1^16A-Flag) were induced with galactose, arrested with αF or noc for 2.5 hours and examined after cycloheximide (CHX) addition as in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005162#pgen.1005162.g001" target="_blank">Fig 1B</a>. Cdc28 is shown as a loading control. B) Quantification of the experiment in [A] is shown. On the left is the quantification from the darker exposure, while right side shows relative abundance quantified from the lighter exposure. Y-axis shows the normalized signal above background normalized to the value at t = 0 in nocodazole arrested Ndd1^wt cells (left) or Ndd1^16A cells (right). C) <i>GAL1p</i>-Ndd1^wt-Flag was induced with galactose in a wildtype or <i>cdc28-as1</i> strain. Cells were then arrested in metaphase with nocodazole for 3.5 hours, with treatment of 1 μM 1-NM-PP1 for the last hour, as noted. Cycloheximide was then added at t = 0 and turnover was followed as in [A]. Cdc28 is shown as a loading control.