Fig. 3.

FTY720 induces programmed cell death in MM cells. a MILL cells were treated with 6 µM FTY720 and the levels of phosphorylated AKT (Ser⁴⁷³), total Bcl-2 and Bax proteins were assessed by immunoblot analysis at indicated time-points. Levels of α-tubulin were detected as loading control. b Activation of PARP and caspase-3 was evaluated in REN cells by immunoblot analysis after treatment with 6 µM FTY720 at indicated time points. Arrows indicate caspase-3 and PARP cleaved forms. c–e Cyt-c and AIF release in PHI cells were assessed by immunoblot analysis. Cells were treated with 6 µM FTY720 or vehicle for 12 and 24 h. Fractionation was performed as described in the “Methods” section. VDAC, lamin and α-tubulin were used as controls of loading and fractions’ purity. Numbers above blotting figures indicate the band densitometric analysis data, relative to loading control. Figures show results from one experiment representative of two independently performed. All immunoblot experiments were repeated with two other MM cell lines with comparable results