ATP Assay

Intracellular ATP was measured using the ATP determination kit (Molecular Probes) according to the manufacturer’s protocol. Mitochondria were isolated as described above and lysed in RIPA buffer. Standard reaction solutions containing 8.9 mL dH2O, 0.5 mL 20× reaction buffer, 0.1 mL 0.1M DTT, 0.5 mL of 10 mM D-luciferin and 2.5 uL firefly luciferase (5 mg/mL) were prepared. 10 μL of the lysed mitochondria was added to 90 μL of the standard reaction solution and the luminescence was immediately read using a luminometer (Femtomaster FB 12 luminometer, Zylux).