MMP expression in <i>Hpse-</i>KO mice.

<p>A. Real-time PCR. RNA was extracted from liver, kidney and mammary gland of <i>wt</i> and <i>Hpse-</i>KO mice and subjected to quantitative real time PCR analysis to evaluate the expression of MMP-2, MMP-9, MMP-14 and MMP-25. The expression level determined for each MMP in the <i>wt</i> tissue (blue) was regarded as 100% and the corresponding expression determined in the <i>Hpse-</i>KO tissue (Purple) are presented as percentage relative to it. Each reaction was repeated 6 times and the mean±SD is indicated. B. Western blot analysis. Liver, kidney and mammary gland tissue extracts, prepared as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005181#s2" target="_blank">Materials and Methods</a>”, were subjected to Western blot analysis using anti-mouse MMP-2 monoclonal antibodies (mA801B; upper panels), anti mouse β-catenin (mAb610154; middle panels), or anti mouse α-tubulin (B-5-1-2; lower panels). Higher protein levels of MMP-2 and β-catenin were detected in samples derived from <i>Hpse-</i>KO vs. <i>wt</i> tissues. C. MMP2 zymography. Serum samples derived from <i>Hpse-</i>KO and <i>wt</i> mice were evaluated for MMP-2 activity. MMP-2 activity was approximately 3 fold higher in plasma samples derived from <i>Hpse-</i>KO vs. <i>wt</i> mice. D. β-catenin immunostaining. Parafin embedded kidney tissue sections were subjected to immunostaining with antibody directed against β-catenin. Increased staining was observed in kidney derived from <i>Hpse-</i>KO vs. <i>wt</i> mice. C. MMP2 zymography. Serum samples derived from <i>Hpse-</i>KO and <i>wt</i> mice were evaluated for MMP-2 activity. MMP-2 activity was approximately 3 fold higher in plasma samples derived from <i>Hpse-</i>KO vs. <i>wt</i> mice.