Figure 1 of
Lin, Mol Vis 2005;
Figure 1. Expression of SOD1 in intact lenses in culture
Fresh lenses from 6-week-old Sprague Dawley rats were immediately incubated with serum-free DMEM (low glucose) for 24 h. Clear lenses were further incubated with human EYFP:SOD1 plasmid DNA at 50 μg/ml for additional 24 h. After that, whole lenses were homogenized and the supernatants were used for detection of expression of SOD1 by western blotting and SOD enzyme activity assay as described in Materials. A: Expression of SOD1 by western blot. Proteins were resolved by SDS-PAGE and immunoblotted with anti-GFP antibody. Expression of EYFP:SOD1 and EYFP was revealed. No signal was detected from lens samples with no plasmid treatment (control). B: Expression of EYFP:SOD1 fusion proteins by western blot using anti-SOD1 antibody. Lower bands show endogenous SOD1 in the lens. C: SOD enzyme activity assay. The supernatants from whole lens extracts were used for SOD enzyme sources to test total SOD enzyme activity. Incubation with SOD1 plasmid DNA causes significant increased lens SOD enzyme activity (asterisk) compared to that in the control (untreated) lenses (control expressed as 100%). Values for three experiments are summarized in a bar graph; the error bars represent the standard error of the mean.