Figure 4. DMF application induces Smac release.
(A) Shown are representative z-stacks of cells treated with DMSO (Veh) or DMF (50 μM) for 8 h. Paraformaldehyde fixed cells were stained for Smac (red), mitochondria (green), Hoechst (blue), and analyzed by confocal microscopy; arrows indicate Smac release; scale bar, 10 μm. (B) Mitochondria were isolated from DMF (50 μM) treated cells as indicated. Cytoplasmic and mitochondrial fractions were analyzed by Western blot. (C) DMSO (Veh), zVad, and caspase-8 inhibitor (C8-I) incubated cells were left untreated or stimulated with DMF (50 μM) for 8 h. Shown are Western blot analyses of isolated mitochondria (Mito). (D) Protein lysates of cells treated as described in (C) were analyzed for expression of cIAP2, cIAP1, XIAP and Actin.