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Fig. 7. Competitive leptin-SEAP binding assay of leptin mutants. We tested the binding of leptin mutants using a competitive leptin-SEAP assay as described in the Materials and Methods section. Wild-type leptin and leptin mutants that bind to CRH2 lead to a decreased binding of leptin-SEAP, measured as decreased alkaline phosphatase activity. The L86 mutants show a decreased competitive binding to CRH2, whereas the L89S and F92S mutations have no effect on binding.





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