Figure 2
From: L166P mutant DJ-1 promotes cell death by dissociating Bax from mitochondrial Bcl-XL
Interactions between Bcl-X L and DJ-1(L166P). (A) The supernatant of E. coli crude extract containing 50 ÎĽg of recombinant Bcl-XL was incubated with 20 ÎĽg of GST, GST-DJ-1 or GST-DJ-1(L166P) coupled to glutathione-agarose beads. The bound proteins were detected by immunoblot analysis. (B) H1299 cells transiently transfected with Flag-DJ-1 or Flag-DJ-1(L166P) were subjected to immunoprecipitation with normal mouse serum or anti-Bcl-XL antibody. (C) HEK293 cells transfected with Myc, DJ-1-Myc or DJ-1(L166P)-Myc along with EGFP-Bcl-XL were subjected to immunocytochemical staining with anti-Myc antibodies (red) and then labeled with DAPI (blue). Bar, 10 ÎĽm. (D) H1299 cells transfected with Flag-DJ-1(L166P) along with EGFP or EGFP-Bcl-XL were treated with or without 80 mJ/cm2 UVB irradiation. Supernatants of the cell lysates were then subjected to immunoprecipitation with anti-GFP antibodies. (E) HEK293 cells transiently transfected with RFP or DJ-1(L166P)-RFP along with EGFP-Bcl-XL were treated with or without 80 mJ/cm2 UVB irradiation, and then the cells were observed with an inverted fluorescence microscope (Olympus, IX71). Bar, 10 ÎĽm.