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HPLC analysis of neurotensin (NT) degradation by B16F10-Nex2 supernatant. Neurotensin (20 μM) was incubated for 1 h at 37°C with recombinant TOP (A), neurolysin (B) or B16F10-Nex2 supernatant (D) in 50 mM Tris-HCl, pH 7.4. The reaction products were separated by HPLC. HPLC profile of melanoma supernatant without NT is shown on panel C. The neurotensin fragments were determined by mass spectrometry.
