Testicular protein extracts were prepared as follows: Testicular tissue (500 mg) was homogenized in ice-cold protein lysis buffer (pH 7.5) containing 50 mM Tris, 250 mM sucrose, 10 mM ATP, 5 mM MgCl 2 , 1 mM DTT, and protease inhibitors (10 g.ml 1 of antipain, aprotinin, leupeptin and pepstatin A, as well as 20 M PMSF)
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Calculating 13 C-labelling enrichment in the detected metabolites The distribution of 13 C-labelling from arginine or proline in the identified metabolite pool was determined by calculating the ratio of 13 C to 12 C in the major mass fragments