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Fig. 8. CNP effects on FGF2-mediated induction and activation of matrix-remodeling molecules. (A) Cells were treated with FGF2 for 4 or 24 hours and the expression of the listed molecules was assayed by RT-PCR. Levels of Gapdh serve as a control for template quantity. (B) Cells were treated with FGF2 and either CNP or the MEK inhibitor U0126 for 4 hours and the amounts of Mmp3, Mmp10, Mmp13, Adamts1 and Adamts5 mRNAs were determined by real-time RT-PCR. Transcript quantities are relative to untreated cells. Mmp2 and Mmp9 were not studied due to the possible nonspecific actions of U0126 during the prolonged cultivation time necessary for their induction. (C,D) Cells were treated with FGF2 and CNP for 72 hours and the conditioned culture media was analyzed for MMP activity (C) by zymography using gelatin (upper figure) or casein (lower figure) as a substrate or (D) for presence of MMP13 by WB. The zymography signal was quantified by densitometry and graphed.
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