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135 Further, PRAS40 and mTOR also exerts negative feedbacks on proximal insulin signaling, PRAS40 knockdown significantly decreases Akt phosphorylation, mTORC1 binds and inhibits INSR by inducing destabilization of IRS1

Supernatants were analyzed by a high-performance liquid chromatography (HPLC) system (The UltiMate 3000 Analytical systems, Thermo Fisher Scientific, USA) according to Zhao et al