p53 induction and Ser15 phosphorylation in B-CLL and in normal cells upon treatment with nutlin-3. Induction of p53 and levels of Ser15 phosphorylated-p53 (P-p53) were assessed by Western analysis. (A) Dose-dependent induction of p53 in normal CD19+ B and B-CLL cells after 24 hours of incubation with increasing doses of nutlin-3. Representative examples of 4 independent experiments are shown. (B) Normal CD19+ B and B-CLL cells were either left untreated or incubated with nutlin-3 (10 μM), or the proteasome inhibitor (Prot Inhib) clasto-lactacystin (25 μM), or fludarabine (10 μM) for the indicated time. Representative examples of 3 independent experiments are shown. (C) Normal PBMCs and BMMCs were either left untreated or incubated with nutlin-3 (10 μM) or fludarabine (10 μM) for 24 hours. Representative examples of 3 BMMC or 5 PBMC independent experiments are shown.
