DIM reduces expression of cell-cycle regulatory proteins.

<p>Following either 12 hr (gray bars) or 24 hr (black bars) treatment with increasing concentrations of DIM, CEM cells were harvested and protein immunoassays were performed for detection of CCND3, CDK4 and CDK6 proteins (three replicate experiments performed). (A) A representative immunoblot is shown for each protein assay. (B) Values shown are average protein expression ± SEM normalized to β-actin, expressed as a percentage difference from time-matched vehicle controls (0.1% DMSO), which were assigned a value of 100%. *, <i>p</i><0.05 or **, <i>p</i><0.01 compared to 0 µM DIM (vehicle control) as determined by one-way ANOVA with Dunnett's post-hoc test for multiple comparisons; overall ANOVA <i>p</i>-values within each time group are indicated in each panel. In some cases where the <i>p</i>-value for the ANOVA was not <0.05, a significant linear trend was evident, as indicated by trend <i>p</i>-values in the figure. Finally, a Student's <i>t-</i>test (###, <i>p</i><0.001) was performed to compare 15 µM DIM to vehicle control for CCND3 expression at 24 hr because high variability observed at the 3.8 µM concentration confounded the ANOVA post-hoc results (overall effect of DIM was significant).