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Louis, MO, USA) were incubated with in vitro -translated proteins (5 l) (TnT Quick Coupled Transcription/Translation Systems, L1170, Promega, Madison, WI, USA) or recombinant proteins (50100 ng) (ProteinOne, Rockville, MD, USA) in a buffer containing 20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 2 mM EDTA, 2 mM DTT, 5% glycerol, and 0.4% NP-40 at 4C overnight, followed by extensive washes with a similar buffer containing 150 mM NaCl

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TMT reporter ion intensities were normalized to the median summed signal intensity across channels, and the data were filtered to retain proteins with at least two distinct peptides

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Suppressed spontaneous and stimulated growth hormone secretion in patients with Cushings disease before and after surgical cure