DIM increases the levels of p21 and p27 proteins and mRNAs in HT-29 cells. Cells were treated with different concentrations of DIM as described in Figure 1. (A) Cell lysates were analyzed by Western blotting with the indicated antibodies. Photographs of the chemiluminescent detection of the blots, which were representative of three independent experiments, are shown. (B) Total RNA was isolated and reverse-transcribed, and PCR was performed with the specific primers for p16, p21, p27, or β-actin. PCR products were separated in an agarose gel with TAE buffer and stained with ethidium bromide. Photographs of the ethidium bromide-stained gels, which are representative of three independent experiments, are shown. The relative abundance of each band to its own β-actin was quantified, and the control levels were set at 100%. The adjusted mean ± SEM (n = 3) of each band is shown above each blot. *Significantly different from 0 μmol/L DIM, P < 0.05.