Fig. 5

Effects of the DNA methylase inhibitor decitabine. U266 cells were stimulated with anti-CD28-loaded particles and cultured without or with 1 µM decitabine for 72 h. A Percentages of cells expressing proteasome subunits, assessed by flow cytometry. B Representative western blot of PI3K/AKT pathway components. C Densitometric analysis of the experiment shown in (B). Band intensities were normalized to β-actin and then to untreated cells (dotted line). D Levels of DNMT3B mRNA and miR29b normalized to those of GAPDH and RNU44, respectively, and expressed as fold change relative to the average value for untreated cells (real-time PCR). E Western blot and densitometric analysis of DNMT3B, with values normalized to β-actin and then to untreated cells. Densitometric and fold change values are mean and SD for 7 independent experiments. *P < .05, **P ≤ .01, Wilcoxon signed-rank test