Figure 2

9AA-induced inhibitory effects on cdk2/cyclin E activity in infected and uninfected cells. (A) ACH2 and CEM cells were treated with various concentrations of 9AA (0.1, 0.5, 1.0 uM) for 24 hrs. Cells were harvested and lysed for immunoprecipitation (IP) with α-Cyc E ab followed by kinase assays. Histone H1 was used as substrate and was added to each reaction tube along with (γ-³²P) ATP (3000 Ci/mmol). Reactions were incubated at 37°C for 30 minutes and stopped by the addition Laemmli buffer. The samples were then separated on a 4–20% Tris-Glycine gel. The gel was dried and exposed to a PhosphorImager cassette and analyzed utilizing Molecular Dynamic's ImageQuant Software. (B) The lysates from (A) were subjected to western blot to evaluate the levels of cdk2 in samples treated with 9AA at different concentrations (0, 0.1, 0.5, 1.0 uM).