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. Author manuscript; available in PMC: 2012 Jun 10.
Published in final edited form as: Mol Cell. 2011 Jun 10;42(5):624–636. doi: 10.1016/j.molcel.2011.03.031

Figure 1. Cdk2 activity is required for human cell proliferation.

Figure 1

(A) The Cdk2 locus was targeted by an rAAV vector containing a neomycin resistance (neoR) cassette bounded by loxP sites and homologous arms, as indicated. The F80G mutation and EcoRI site used for genotyping were introduced in exon 3.

(B) RPE-hTERT cells with varying amounts of total and AS Cdk2, quantified by immunoblot.

(C) HCT116 and RPE-hTERT whole-cell extract proteins were labeled by endogenously expressed Cdk2as with [γ-32P]N6-(benzyl)-ATP.

(D) RPE-hTERT cells of the indicated genotype were grown for 7 d in the presence of increasing amounts of 3-MB-PP1. Cells were fixed and stained with crystal violet solution.

(E) RPE-hTERT cells of the indicated genotype were treated with DMSO or 1 μM 3-MB-PP1 for 48 hr prior to seeding. Cells were maintained in DMSO or, if pre-treated with 1 μM 3-MB-PP1, switched to 10 μM 3-MB-PP1, and cell number was determined every 24 hr. Each data point is the mean +/− SEM of 3 independent plates of cells.