Figure 7
Effect of PAI-1 mutants and uPA-PAI-1 complex on matrix formation. A – MG-63 cells were incubated overnight on vitronectin-coated wells in serum-free media. Cells were then pretreated with 1 μg/ml (20 nM) wild-type PAI-1 or PAI-1 mutants (Q123K or PAI-1R) for 20 min in the presence of 125I-fibronectin and 50 μM peptide was added for 6 hours. Cell layers were extracted in 1% DOC, and 125I-fibronectin that was incorporated into the detergent-insoluble matrix was recovered by centrifugation and measured by gamma scintillation. B – Cells were pretreated with 20 nM wild-type PAI-1 or uPA-PAI-1 complex for 20 min in the presence of 125I-fibronectin and 50 μM peptide was added for 6 hours. Cell layers were processed as described in A.