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. 2013 May 13;110(22):8954–8959. doi: 10.1073/pnas.1302927110

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Fig. 1. — Cdk2^AF/AF cells display early S-phase entry and accelerated cyclin E turnover. (A) Lysates were harvested from Cdk2^+/+ and Cdk2^AF/AF cells. Phospho-Y15 Cdk2 and total Cdk2 abundance were measured by Western blotting. (B) Cdk2^+/+ and Cdk2^AF/AF cells were arrested in G₀/G₁ by serum/leucine starvation and released into media containing nocodazole. Samples were harvested as indicated, and DNA content was analyzed by flow cytometry. (C) Cyclin E-Cdk2 abundance and kinase activity were assayed in the samples harvested in B. (D) Cyclin E half-life was determined by pulse–chase analysis beginning 13 h after serum/leucine release. (E) The indicated cell lines were treated with HU as indicated and assayed for cyclin E and Cdk2 levels. (C and E) PP2A-loading control. (F) Cyclin E turnover was measured in HU-arrested Cdk2^+/+ and Cdk2^AF/AF cells by pulse–chase analysis.