2.5. Acetyl-CoA and malonyl-CoA levels

Cells from a 6-well plate were extracted with 110 μl of 5% perchloric acid (Sigma–Aldrich). The samples were then centrifuged at 15,000×g (10 min at 4 °C). The supernatant was transferred to a vial and 3 μl was loaded onto a 6495 Triple Quad LC/MS (Agilent, Santa Clara, CA, USA) equipped with a BEH amide column (2.1 mm × 100 mm and 1.7 μm particles, Waters). The transition to quantify Acetyl-CoA is 810 -> 303. The abundance of acetyl-CoA was expressed as peak areas on a mass spectrometer and calculated with Agilent MassHunter Workstation Software. The level of malonyl-CoA was measured by a human malonyl coenzyme A ELISA Kit from CUSABIO as instructed (Cusabio, Houston, TX, USA).