Fig. 1.

Stable over-expression of OGC specifically increases the mitochondrial glutathione (GSH) pool. (a) Whole cell lysates were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotted against V5, OGC, neuron-specific β3 tubulin, DIC, γ-glutamylcysteine ligase (GCL) catalytic subunit, and actin. OGC stable cell lines are indicated as OGC 3 and OGC 6; parental NSC34 cells are indicated as wild type (WT). NS, non-specific band. (b) Cells were fractionated by differential centrifugation and total mitochondrial GSH content was then measured using a DTNB colorimetric assay. Data are represented as a percentage of WT GSH measurements. *p < 0.05 versus WT mitochondrial GSH; **p < 0.01 versus WT mitochondrial GSH in an unpaired Student’s t-test, n = 7, error bars indicate SE. (c) Cells were fractionated by differential centrifugation and total cytosolic GSH content was then measured using a DTNB colorimetric assay. Data are represented as a percentage of WT cytosolic GSH measurements. n = 7, error bars indicate SE. (d) Cellular fractions were resolved using SDS–PAGE and immunoblotted against Cox-IV and α-tubulin (α-Tub).