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. 2005 Jun 27;102(27):9691–9696. doi: 10.1073/pnas.0409635102

Fig. 4.

Fig. 4.

DJ-1 protects against Daxx/ASK1-induced cell death and inhibits ASK1 activation. (A) Wild-type but not L166P mutant DJ-1 protects against Daxx/ASK1-induced cell death. SH-SY5Y cells were transfected with the indicated expression vectors, and 24 h later, cell death was quantified by using EthD-1. Values shown are means ± SEM for triplicates. *, ANOVA P < 0.001. (B) DJ-1 blocks Daxx-induced ASK1 activation. COS-7 cells were transfected as indicated, followed by immunoprecipitation with anti-Myc. In vitro kinase assay was performed with the immunoprecipitation complex. Expressions of Myc-ASK1, HA-Daxx, and FLAG-DJ-1 were checked by Western blot analysis. W, wild-type DJ-1; M, L166P mutant. (C) DJ-1 inhibits H2O2-induced ASK1 activation. COS-7 cells were transfected as indicated and challenged with 0.2 mM H2O2 for 1 h. (D) DJ-1 inhibits the interaction between ASK1 and Daxx. COS-7 cells were transfected with 1 μg of the indicated plasmids except for L166P (3 μg). (E) Reduced expression of DJ-1 exaggerates H2O2-induced ASK1 activation. SH-SY5Y cells were transiently transfected with 100 μM siRNA against DJ-1 or mutant siRNA for 48 h, followed by 0.2 mM H2O2 treatment for 30 min.