Figure 1.

Effect of Grx1 on SOD1 solubility in neuronal cells. (A) Evaluation of enzymatic activity of Grx1 expressed in μmol/NADH/min/mg protein in total extracts from both NSC-34 and SH-SY5Y cells either untransfected or overexpressing Grx1 (upper panel) and the western blot analysis of Grx1 expression levels in the same cell lines (lower panel). Results represent mean ± SD from three independent experiments. Values significantly different from relative controls are indicated with an asterisk when P < 0.01. (B) NSC-34 and SH-SY5Y cells, either untransfected or overexpressing Grx1, were infected with an adenoviral vector leading the transient expression of wild-type SOD1 or G93A-SOD1. The soluble and insoluble protein fractions were collected from cellular lysates and equal amounts of proteins were subjected to a denaturing PAGE as described in Materials and Methods, followed by western blot analysis with an anti-SOD1 antibody. Endogenous mouse SOD1 (mSOD1) serves as an internal loading control. (C) SH-SY5Y cells, either untransfected or overexpressing Grx1, were infected with an adenoviral vector leading the transient expression of wild-type SOD1 or G93A-SOD1. Cells were sub-fractionated on gradients; the soluble and insoluble protein fractions were collected from mitochondrial fractions and subjected to a denaturing PAGE as described in Materials and Methods, followed by western blot analysis with an anti-SOD1 antibody. The blot shown is representative of three independent experiments.