MIP-1α phosphorylates FKHR, downstream target of AKT/PKB pathway in MM cells.

(A) MM1.S cells and (B) pat MM cells were incubated with MIP-1α (100 ng/mL) for 5 and 15 minutes. Pretreatment with PI3-K inhibitor wortmannin (1 μM) or LY 294002 (50 μM) was performed for 1 hour prior to cytokine treatment. Cells were lysed, electrophoresed, and immunoblotted with antiphospho-FKHR (Ser256) antibody. This mab is cross-reactive with AFX, which is constitutively expressed in MM1.S and pat MM cells and served as a loading control (ctr).