2D zymography reveals gelatinase isoforms in mouse brains after focal cerebral ischemia.

<p>Mice were sacrificed 24 h after filament-induced MCAo in mice. (<b>A</b>) 1D zymography revealed increases in proMMP-9 and act.MMP-9 levels in the ischemic cortex compared with the contralateral cortex. Under these experimental conditions, proMMP-2 was not altered. Densitometry analysis of intensity of gelatinolytic bands represented proMMP-9 and act.MMP-9, n = 4, *, <i>p</i><0.001, comparing ischemic to contralateral cortex by one-tailed, unpaired Student’s <i>t</i>-test; data are expressed as mean values ± SEM. (<b>B</b>) Brain lysate was incubated with gelatin 4B and applied on 2D gels. In contralateral hemispheres, proMMP-9 was identified as a 105-kDa single spot with pI value between 3 and 4, and proMMP-2 as a 65-kDa single spot with pI value between 4 and 5. In ischemic hemispheres, two streaks of pI values ranging from 5.5 to 8 with the molecular weights at 105 and 95 kDa were identified as proMMP-9 and act.MMP-9, respectively. Ischemic brain lysate was applied on the left side of the gel for comparison. These zymograms are representative results from 3 independent experiments.