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Effects of chronic ET-1 treatment on insulin-stimulated Akt phosphorylation. Serum-starved 3T3-L1 adipocytes were incubated in the absence or presence of insulin (100 ng/ml) for 10 minutes, after treatment with or without ET-1 (10 nM) for 24 hours. Whole-cell lysates were subjected to SDS-PAGE and immunoblotted with phospho-specific Akt (pAKT) serine 473 (top panel)/threonine 308 (middle panel), or anti-Akt-1 antibody (lower panel). These experiments were repeated twice.
