Protein extracts were generated by homogenization in an ice-cold M-PER protein extraction reagent (Thermo Fisher Scientific, Rockford, IL) containing a protease inhibitor cocktail (Complete Mini and phosphoSTOP, Roche Diagnostics, Basel, Switzerland). The protein content of the lysate was quantitated using Pierce BCA Assay Reagent (Thermo Fisher Scientific). Primary antibodies for Western blot analysis were FOXO1 (C29H4) (#2880, Cell Signaling, Boston, MA, USA) and α-tubulin (#T6074, Sigma-Aldrich, St. Louis, MO, USA). A horseradish peroxidase (HRP)–conjugated polyclonal goat anti-rabbit (#P0448, DAKO, Glostrup, Denmark) was used as secondary antibody for FOXO1, and for the anti–α-tubulin antibody, an HRP-conjugated goat anti-mouse (#P0447, DAKO) was used. Enhanced chemiluminescence reagents (Pierce, Thermo Fisher Scientific) were used for detection. Gels were imaged and analyzed with Image Lab Software (Bio-Rad Laboratories Inc., CA, USA)

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