The molecular mechanisms for cell-adhesion–mediated Skp2 and Cdh1 changes. (A,B) Effect of cell adhesion on Skp2 degradation. Jeko-1 cells were treated with cycloheximide (100 μM) with and without HS-5 cells adhesion for the indicated time periods. At the times indicated, cells were lysed, and Skp2 protein levels were determined by Western blot analysis (A) and quantified (from 3 independent experiments) using densitometry. The data shown (B) represent mean values and standard deviations of the average percentage of Skp2, compared with the amount of Skp2 at time 0 for each treatment group. (C,D) Cell-adhesion–induced changes of Skp2 and CDH1 mRNA expressions measured by real-time qRT-PCR. Fold values were obtained by externally standardizing against identical amplifications in Jeko-1 and SUDH-10 cells in suspension versus HS-5 adhesion and by internally standardizing against GAPDH in each cell line. The mean values and standard deviations from 3 independent experiments are shown. Student t test was used for statistical analysis. *P < .05.