Cocktail inhibitor

All cell culture media (Dulbecco’s modified Eagle’s medium, phosphate-buffered saline, and penicillin-streptomycin), sodium palmitate, sodium pyruvate, 2-deoxy-D-glucose were purchased from Sigma Aldrich (St. Louis, MO, USA). Foetal bovine serum (FBS), horse serum, cell extraction buffer and cocktail inhibitor were obtained from Life Technologies (Warrington, UK). Tritiated glucose, D-[3-3H] was obtained from Perkin Elmer (Boston, MA, USA). Mitochondrial ATP monitoring reagent was purchased from Biothema (Handen, Sweden). Protein assay reagents were purchased from BioRad (Copenhagen, Denmark). Anti-PDK4 (ab214938), anti-PPAR alpha (ab24509), anti-PPAR delta (ab23673), goat anti-Mouse (IRDye^® 800CW) (ab216772), goat Anti-Rabbit (IRDye^® 680RD) (ab216777) and anti-alpha actin (ab28052) antibodies were purchased from Abcam (Cambridge, UK). Anti-Phospho-FoxO1 (Ser256) (#9461) and Anti-FoxO1 (#2880) antibodies were purchased from Cell Signaling Technology (London, UK). Levels of cytosolic and nuclear fractions of p- and t-total FOXO1 were identified using the same antibodies that were used for total cellular lysate measurements but were applied to isolated cellular fractions using a nuclear protein extraction kit (Active Motif Europe, La Hulpe, Belgium).

Cell culture media (Dulbecco’s modified Eagle’s medium, phosphate-buffered saline and penicillin-streptomycin), sodium pyruvate, sodium palmitate and 2-deoxy-D-glucose were purchased from Sigma Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), horse serum, cell extraction buffer and cocktail inhibitor were acquired from Life Technologies (Inchinnan, UK). Tritiated glucose, D-[3-3H], was obtained from Perkin Elmer (Boston, MA, USA). Mitochondrial ATP monitoring reagent was acquired from Biothema (Handen, Sweden). Protein assay reagents were purchased from BioRad (Copenhagen, Denmark). Anti-PDK4 (ab214938), anti-PPAR alpha (ab24509), anti-PPAR delta (ab23673), goat anti-Mouse (IRDye^® 800CW) (ab216772), goat Anti-Rabbit (IRDye^® 680RD) (ab216777) and anti-skeletal muscle alpha-actin (ab28052) antibodies were acquired from Abcam (Cambridge, UK). Anti-Phospho-FoxO1 (Ser256) (#9461) and Anti-FoxO1 (#2880) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).