Dimethyl Fumarate Induces Glutathione Recycling by Upregulation of Glutathione Reductase

Figure 2

Knockdown of GSR boosts the protective effect of DMF by inducing a synergistic set of antioxidant response genes. (a) Cells were transfected with siRNAs against GSR (siGSR) or luciferase (siCtr) 24 h before addition of 10 μM DMF for an additional 24 h. The same amount of protein lysates was blotted and stained with antibodies against GSR or actin as loading control. The molecular weight is indicated. The bar graphs represent the means ± SEM normalized to vehicle of 3 different blots. (b) DMF was added 24 h after transfection of the indicated siRNAs and 10 mM glutamate 24 h after DMF. Viability was quantified by CTB assays again 24 h later. The bar graphs represent the mean ± SEM of three experiments done in triplicate. (c) DMF treatment or siGSR transfection induces mRNA expression of a synergistic set of transcripts involved in the antioxidant response. Cells were treated for 24 h with 10 μM DMF or vehicle or prepared 48 h after transfection with siGSR and siCtrl and mRNA quantitated by real-time PCR using β-actin and hprt as endogenous controls. , two-way ANOVA, and Tukey’s post hoc test.

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