Fig. 4

Prkn expression alters glutathione metabolism in murine brain, including the activity of glutathione reductase. a Tietze method-based quantification of reduced glutathione (GSH), oxidized glutathione (GSSG), the ratio of GSH:GSSG, and total glutathione (GSH + 2GSSG) in mouse brain homogenates of two genotypes, as indicated. b Schema of GSH synthesis pathway, where glutamate cysteine ligase (GCL) is the rate limiting enzyme; GS = glutathione synthase. c Relative GCLC, GCLM and Dj-1 mRNA levels in the brains of 6 month-old mice (–C and –M denote catalytic and modifying subunits respectively; n = 3 mice/genotype). d HPLC-based quantification of GSH, GSSG, the ratio of GSH:GSSG, and the total glutathione pool (GSH + 2GSSG) in brains of 6 month-old mice, as indicated. e, f Western blot analysis of glutathione reductase (GR) protein levels brains from 6 month-old mice, as separated by SDS/PAGE (reducing conditions) using Ponceau S as the loading control, and its quantification by densitometry (n = 6/genotype) in (f). g GR activity in freshly prepared brain homogenates of 6-month-old mice (n = 3/genotype), as indicated. Significance was determined using unpaired Student T-test (a, c, d, f, g) where * represents p ≤ 0.05 and **p ≤ 0.01