Figure 6

Effects of compound-23 on excess oxidation and dimer formation of DJ-1. (A) SH-SY5Y cells were incubated with 1 μM compound-23 or compound-B for 20 hours and then treated with various amounts of H₂O₂ for 15 min. Oxidation of DJ-1 in cells were analyzed by using isoelectric focusing phoresis followed by Western blotting with an anti-DJ-1 antibody (upper panel). Intensity of bands was quantified and ratio of oxidized DJ-1 to total DJ-1 is shown (lower panel). (B) SH-SY5Y cells were incubated with 1 μM compound-23 or compound-B for 20 hours and then treated with various amounts of H₂O₂ for 3 hours. Cells were then treated with 0.5 mM DSS or DMSO for 30 min, and proteins extracted from cells were analyzed by Western blotting with an anti-DJ-1 antibody.