Effect of Cand1 dose on the stability of the SCF substrate Rum1p in S. pombe.

A: S. pombe cells stably overexpressing the indicated versions of Cand1 were treated with 100 μg/ml cycloheximide (CHX) for the indicated periods of time and analyzed for the expression of Rum1p, Cul1p, and Cand1 by immunoblotting. Cdc2p signals are shown for reference. Representative results of two independent experiments. B: Immunoblotting signals for Rum1p and Cdc2p were quantified using Image Studio Lite, and Rum1p intensities normalized to Cdc2p were plotted on a log scale. C:Exponential decay lines fitted through the data points in (B) were used to calculate protein half-lives in strains overexpressing the indicated Cand1 proteins. Results are averages of two independent experiments. Error bars represent standard deviations.