Supplementary Material
JCS009530 Supplementary Material
Files in this Data Supplement:
- Supplemental Figure S1 (Adobe PDF) -
Fig. S1. Measurement of stoichiometry of CDK2 phosphorylation. (A) The recombinant CDK2 was phosphorylated in the presence of either active Akt or inactive Akt using γ-32P ATP. Silver-stained CDK2 protein bands were excised from the gel and the amount of incorporated 32P was quantitated by scintillation counting and plotted against the concentration of the protein used in the kinase assay. The data represent the mean of ± s.e.m. of three independent experiments. (B) CDK2 wild-type (WT) proteins or those mutated at T39, T160 or both were used in a kinase assay with a recombinant active Akt, and the stoichiometry of CDK2 phosphorylation was determined as described in A. The data represent the mean of ± s.e.m. of three independent experiments.
- Supplemental Figure S2 (Adobe PDF) -
Fig. S2. 3T3 fibroblasts transfected with CDK2 T39A mutants were synchronized in different indicated phases of the cell cycle and the localization of the CDK2 mutant was detected by staining with HA antibody followed by Cy3-conjugated secondary antibody. DAPI was used to counterstain the nucleus. The images were acquired and analyzed using an Olympus-IX81 multilaser confocal microscope.