Figure 5
From: L166P mutant DJ-1 promotes cell death by dissociating Bax from mitochondrial Bcl-XL
Promotion of cell death by DJ-1(L166P)-influenced Bcl-X L /Bax heterodimerization. (A) Cell lysates of H1299 cells stably expressing sh-NC or sh-DJ-1 were subjected to immunoblot analysis. (B) The relative ratios of DJ-1 to α-Tubulin in (A) were analyzed using densitometry (mean ± S.E.M., n = 3; ***, p <0.001; by one-way ANOVA). (C) H1299 cells stably expressing sh-DJ-1 were transfected with Flag, Flag-DJ-1(s) or Flag-DJ-1(L166P)(s) and treated with or without 80 mJ/cm2 UVB irradiation. The cells were then subjected to subcellular fractionation. (D) The relative level of Bax in (C) were analyzed using densitometry (mean ± S.E.M., n = 3; *, p <0.05; **, p <0.01; by one-way ANOVA). (E) The relative level of Bcl-XL in mitochondria in (C) were analyzed using densitometry (mean ± S.E.M., n = 3; *, p <0.05; by one-way ANOVA). (F-G) H1299 cells stably expressing sh-DJ-1 transfected with si-NC or si-Bcl-XL along with Flag, Flag-DJ-1(s) or Flag-DJ-1(L166P)(s) were treated with or without 80 mJ/cm2 UVB irradiation. Cell lysates were then subjected to immunoblot analysis (F) or the cells were analyzed with the MTT assay (G) (mean ± S.E.M., n = 3; *, p <0.05; ns, no statistical significance by one-way ANOVA).