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Competition binding assay of agrin mutants. Myotubes grown in fusion medium on 35-mm dishes were incubated with 5 nm 125I-AgG3z8 plus a mutant protein. Ligand binding in the absence of the competitor was normalized to 100%. The amount of bound competing ligand was calculated from the reduction in 125I-AgG3z8. KI values were calculated from the IC50 concentration of competing ligands by the equation of Cheng and Prusoff (46) and presented in Table 1.
