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Phosphorylation of FoxO1 acetylation site mutants. HEK-293 cells were transfected with FLAG-tagged T24A, T24A-KQ, or T24A-KR FoxO1 and treated with different concentrations of insulin. Phosphorylation of FoxO1 and Akt was determined by Western blotting with anti-phospho-S253 FoxO1 (p-S253), phospho-T24 FoxO1 (p-T24), and phospho-Akt (T308) antibodies. Tubulin was used as the gel loading control.
