Untreated (–) and treated (+) 293T whole cell extracts (30 μg) were separated by 12% SDS-PAGE, and the membrane was blotted with PRAS40 (phospho Ser183) antibody [HL3189] (GTX640828) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.