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. 1997 May 5;137(3):779–791. doi: 10.1083/jcb.137.3.779

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Binding of His-pro-uPA^wt and His-pro-uPA^138E/303E to vitronectin in the absence and in the presence of uPAR. (A) 1.25 μg, 2.5 μg, and 5 μg of urea-denatured vitronectin (lanes 3, 4, 5 and 8, 9, 10, respectively), or 5 μg of fibronectin (lanes 1 and 6) or collagen (lanes 2 and 7) were separated onto 7.5% SDS-PAGE under reducing conditions, blotted to a PVDF membrane, and probed with 2.5 × 10⁶ cpm of ¹²⁵I-labeled His-pro-uPA^wt (lanes 1–5) or His-pro-uPA^138E/303E (6–10) in a total volume of 2.5 ml. The filter was dried and exposed to an autoradiographic film. (B) 50,000 cpm of ¹²⁵I-vitronectin were incubated with 10⁶ primed U937 cells, in the absence or in the presence of 10 nM recombinant prouPA, His-pro-uPA^wt, or His-pro-uPA^138E/303E. Specific binding of ¹²⁵I-vitronectin, calculated by subtracting the background obtained in the presence of 100 nM vitronectin, is reported in the figure.