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All chromatography buffers were prepared in Milli-Q Ultrapure water (MilliporeSigma, Burlington, MA) with 1015 mM Tris-HCl (Thermo Fisher Scientific) and 0.005% Polysorbate 80 (PS80) (Croda, Snaith, United Kingdom), and the indicated sodium chloride (NaCl) (Teknova, Hollister, CA) concentration and pH were adjusted with either 50% sodium hydroxide (NaOH) (Thermo Fisher Scientific) or 37% hydrochloric acid (Thermo Fisher Scientific)

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However, due to the limitation of resolution of the density map, subtle structural variations of three arms of ibutamoren are possible