Growth factors and coculture with BMSCs do not protect against perifosine-induced WM cell cytotoxicity. (A) BCWM.1 cells were cultured with control media and with perifosine (5 to 20 μM) for 48 hours in the presence or absence of BMSCs. Cell proliferation was assessed using the [³H]-thymidine uptake assay. (B) BCWM.1 cells were cultured with either perifosine (10 μM) alone and in presence of BMSCs for 6 hours. Whole cell lysates were subjected to Western blotting using anti–p-Akt, -Akt and α-tubulin antibodies. (C) BCWM.1 cells were cultured with perifosine (5 to 20 μM) in the absence and presence of IL-6 (25 ng/mL) or IGF-1 (50 ng/mL) for 48 hours. Cytotoxicity was assessed by the MTT assay. All data represent mean (± SD) of triplicate experiments (A,C). (D,E) BCWM.1 cells were cultured with perifosine (10 μM) for 6 hours in the absence and presence of (D) IGF-1 (50 ng/mL) or (E) IL-6 (25 ng/mL) for the last 10 minutes. Whole cell lysates were subjected to Western blotting using anti–p-Akt ser473, -Akt and α-tubulin antibodies.