Figure 3

MIRA-1 induced time and dose-dependent apoptosis in MM cells. MIRA-1 treatment results in upregulation of Puma and Bax and downregulation of Mcl-1 and c-Myc in a time- and dose-dependent manner with associated activation of caspase-3 and PARP. (A) Total levels of the indicated proteins were evaluated by western blot analysis in the indicated cell lines after treatment with 10 μmol l⁻¹ (MM.1S) or 20 μmol l⁻¹ (LP1) cells for up to 24 h. (B) LP1 cells were treated with indicated concentrations of MIRA-1 for 8 h. Total cell lysates were prepared and analysed by western blot for the expression of the indicated protein. (C and D) p53 siRNA-transfected MM.1S and 8226 cells were further treated with 10 or 20 μmol l⁻¹ MIRA-1, respectively, for 48 h and analysed for apoptosis induction by FCM; NS, not significant.