Core control principles of the eukaryotic cell cycle - Nature

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Core control principles of the eukaryotic cell cycle - Nature
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Nature research paper: Work in fission yeast unites two opposing views of cell cycle control

. Unless otherwise stated, all experiments were conducted in yeast extract medium supplemented with adenine, leucine, histidine and uracil to a final concentration of 0.15 g l. Cells were grown at 25 °C unless stated otherwise. All experiments were performed with cells in exponential growth, defined as 2.5–10 × 10. To induce expression from the tetracycline-dependent promoter, anhydrotetracycline hydrochloride dissolved in DMSO was added to a final concentration of 0.3125 μg ml.

To check for nuclear enrichment of synCut3–mCherry and cyclin–Cdk1–sfGFP, the mean pixel value of a circle encompassing the nucleus was compared to the mean pixel value of a circle of equal area drawn in the cytoplasm. If the nuclear mean value was 1.5 times the cytoplasmic value or greater, this was classed as nuclear enrichment. Scoring for nuclear enrichment indices, cell cycle progression indices and spindle formation indices was conducted on 100 cells per time point.

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