Fig. 7.

GH-induced phosphorylation of ACLY, PRAS40, and raptor is inhibited by wortmannin. A, 3T3–F442A preadipocytes were pretreated with 100 nm wortmannin or vehicle for 30 min and then treated with GH (500 ng/ml) for 30 min. Whole-cell lysates were immunoblotted with αpS455-ACLY, αACLY, αpT247-PRAS40, αPRAS40, αS473-Akt, αpS235/236-S6 kinase, or αS6 kinase as indicated (n = 3). B, 3T3–F442A preadipocytes were pretreated as indicated with vehicle, 100 nm wortmannin, and/or 500 nm CL-1040, for 30 min and then treated with GH (500 ng/ml) for 15 min. Clarified cell lysates were immunoblotted with αpS863-raptor, α-raptor, αpErk1/2, αErk1/2, αpS473-Akt, or αAkt (n = 3).