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

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Fig. 2. — Cdk2 inhibitory phosphorylation is required for recovery from replication stress. (A) Cdk2^+/+ or Cdk2^AF/AF cells were arrested in HU and released into media containing nocodazole. Samples were harvested as indicated, and DNA content was analyzed by flow cytometry. (B) Same as in A, but cells were arrested in Aphidicolin. (C) Experimental setup for D and E: On day 0 cells were seeded; on day 1, HU (D) or Aphidicolin (E) were added and were removed on day 2. Proliferation was assayed 3 d later by using Alamar Blue. Percent proliferation is calculated as percentage of proliferation relative to untreated cells. (F) Annexin V staining of Cdk2^+/+ and Cdk2^AF/AF cells treated with HU or staurosporine for 18 h. (STS, positive control). (G) Cdk2^+/+ and Cdk2^AF/AF cells were treated as in C and assayed for senescence-associated β-galactosidase activity 3 d after HU removal. Error bars indicate SD from three independent data points.