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Fig. 5. TSP-1-induced activation of Erk2. (A) Western blot showing the effect of TSP-1 alone and in the presence of RGD peptide (0.1 mM), function-blocking ß1- integrin antibody (50 µg/ml), heparin (100 µg/ml), function-blocking
vß3 antibody (50 µg/ml) and function-blocking
3ß1 antibody (20 µg/ml) on the activity of Erk. Total Erk2 was immunoprecipitated from whole-cell lysates prior to western blotting with a monoclonal anti-phospho Erk1/2 antibody (0.5 µg/ml, 1 hour). The western blot shown is representative of n=3-10 separate cell strains. (B) The histogram shows the mean activation of Erk2 as determined by densitometric analysis of the dual-phosphorylated kinase. Dual phosphorylation of Erk2 in the TSP-1-stimulated cells was taken to be 100% and other data are normalised with respect to this value. Data represent the mean±s.e.m. of all 3-10 cell strains.