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Proc Soc Exp Biol Med (2000) 223(4):34451.10.1111/j.1525-1373.2000.22349.x 304 SjogrenKLiuJLBladKSkrticSVidalOWalleniusVet alLiver-derived insulin-like growth factor I (IGF-I) is the principal source of IGF-I in blood but is not required for postnatal body growth in mice

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Immunoblot analysis Whole cell lysates were prepared using radio-immunoprecipitation assay (RIPA) lysis buffer (pH 7.8) containing 25 mM Tris-HCl, 120 mM NaCl, 5 mM EDTA and 0.5% NP-40 supplemented with 100 g/mL phenylcmethylsulfonyl fluoride, 200 mM sodium orthovanadate, 0.5 M NaF, protease inhibitor cocktail sets III and IV and phosphatase inhibitor cocktails 2 and 3 (Sigma-Aldrich)