S-Nitrosoglutathione Reductase Plays Opposite Roles in SH-SY5Y Models of Parkinson’s Disease and Amyotrophic Lateral Sclerosis

Figure 4

DJ-1 and AKT in siGSNOR cells and GSNOR-KO brains. Western blot analyses of DJ-1, as well as basal and phosphoactive AKT in total extracts obtained from (a) siScr and siGSNOR SH-SY5Y cells or (b) GSNOR-KO and WT brains. Densitometric analyses of DJ-1 and phospho-AKT are shown on the top of the corresponding Western blot normalized to actin and basal AKT, respectively. (c) Biotin switch assay followed by pull-down with streptavidin and revealed with anti-DJ-1 antibody, in total extracts obtained from GSNOR-KO and WT brains. Western blot analysis indicates that DJ-1 was S-nitrosylated (present in the pull-down). Western blots shown are representative of at least independent experiments that gave similar results. Actin or GAPDH were selected as loading controls. (d) Direct cell count upon Trypan blue staining of siScr and siGSNOR SH-SY5Y cells transfected or not with siRNA against DJ-1 (siDJ-1) and treated for 24 h with 2.5 mM MPP⁺. Graphs shown represent the mean of data ± SD of independent experiments, and .

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