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Proteins were extracted in RIPA buffer (s25 mM tris-HCl (pH 7.6), 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% SDS) with a protease inhibitor cocktail (Roche Diagnostics). Total purified proteins were incubated with 5 mg of antibody specific for IGFBP-3 (Santa Cruz Biotechnology) to form immunocomplexes, which were precipitated with protein A-Sepharose (GE Healthcare). Finally, immunocomplexes were subjected to Western blotting.
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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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