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Fig. 4. Fibronectin rescues cell attachment to human platelet osteonectin after fibronectin (FN) is immunoprecipitated from the human platelet osteonectin (OSN). (A) Fibronectin was immunoprecipitated from human platelet osteonectin (OSN+IP) and cell attachment was assessed as described in Fig. 1. After addition of fibronectin back to the fibronectin-depleted human platelet osteonectin (OSN+IP+FN), cell attachment was restored. The fibronectin added (0.1 µg ml^–1) was 1% of the amount of human platelet osteonectin (10 µg ml^–1) as determined in Fig. 3 to be the amount of fibronectin present in human platelet osteonectin. Immunoprecipitation with control IgG (OSN+IP con IgG) did not alter the level of cell attachment to human platelet osteonectin. Results (mean adhesion ± standard deviation) are shown for MDA-231 breast carcinoma cells. ***P<0.0001 significant difference from OSN tested by Bonferonni's multiple comparison post-test. (B) ELISA results for levels of fibronectin and osteonectin for mixtures used in A. This assay was not linear after absorbance of 0.4 OD and therefore this assay was used to determine the presence or absence of proteins. Minus sign (–) <0.1 OD, one plus sign (+) 0.1-0.4 OD, two plus signs (++) >0.4 OD; OD was measured at 450 nm. Each sample was measured in triplicate. Data are representative of three assays.

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