Figure 5. Small molecule-induced re-distribution of Apaf-1 to the cytosol is ROS dependent.
a. Raji cells were pre-incubated with Catalase (3000U/ml) followed by incubation with C1 (50μg/ml) or LY30 (25μM) for 1hr. Intracellular ROS was assessed using the fluorescent probe DCHF-DA as described in Materials and Methods. b. Lipid raft fractions were purified from the same cells and Apaf-1 expression was checked in fractions 2, 3, 4 and 5 by Western blotting. Flotillin was used as a control for lipid raft resident protein. c. Raji cells were pre-incubated with catalase (3000U/ml) followed by exposure to C1 (25, 50μg/ml) or LY30 (25, 50μM) for 15, 30 and 60 min. Lipid peroxidation was assessed using the fluorescence based assay using Cis-perinaric acid as a probe. *, **, #, ## p < 0.05