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Shc, PRAS40, and raptor are phosphorylated in response to GH. 3T3–F442A preadipocytes were treated (A and C) with GH [500 ng/ml (23 nm)], or (B) with GH (100 ng/ml) for the times indicated. Whole-cell lysates were immunoblotted with αpY423-Shc, αShc, αpS184-PRAS40, αpT247-PRAS40, and αPRAS40 as indicated. Clarified cell lysates were immunoblotted with αpS863-raptor or α-raptor as indicated. The results are representative of three separate experiments. ns, Nonspecific.
