Effect of MVC treatment on MMP-2 and MMP-9 levels in astrocyte culture supernatants.

<p>Primary astrocytes (1×10⁵ cells/ml), incubated in serum-free DMEM, were treated with MVC at the indicated concentrations in the presence of PMA (100 nM) (A) or LPS (10 µg/ml) (B). Negative and positive control supernatants were obtained from unstimulated and untreated astrocytes in serum-free DMEM (control) and cells stimulated with LPS or PMA, respectively. After 24 h of incubation equal amounts of cell culture supernatants were subjected to gelatine zymography as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0028499#s2" target="_blank">Methods</a> section. Representative gels are reported in (A) and (B). MMP-2 and MMP-9 were identified by their apparent molecular mass of 67 and 92 kDa, respectively, using pre-stained molecular weight markers (Bio-Rad). Histograms represent results, expressed as mean±SD, after scanning densitometry and computerized analysis of gels, from at least three independent experiments with different cell populations. Asterisk represents values statistically different from positive control (PMA-treated astrocytes) (One-way Anova followed by Student-Newman-Keuls post hoc test; * = p<0.05).