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The immunoblots were visualized using ECL Western blotting substrate (Pierce Biotechnology) on a Chemidoc XRS imager (Bio Rad) or using Dylight 800-conjugated secondary antibodies (Thermo Fisher Scientific, Rockford, IL) on an Odyssey Imaging Systems (LI-COR Biosciences, Lincoln, NE)

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Here, we prepare a series of Tri-Click ligands incorporating systematic cyclic and acyclic N-, O-, and S-donors and evaluate their AMN activities