a dedicated jugular catheter and arterial blood samples (75 μL) were collected at 2, 5, 10, 15, 20, and 25 min. Then at 30 min an albumin-palmitate-fatty acid tracer complex was infused via another dedicated jugular catheter at 230 μL/min for a total of 4 min as described in [20]. Two fatty acid tracers were included; (1) the partially-metabolizable analogue [9,10-3H]-(R)-2-bromopalmitate (3H-R-BrP, ~5 × 107 dpm, synthesized and purified as described in [21]) and (2) [U-14C]palmitic acid (14C-P, ~2.5 × 107 dpm; Amersham, Solna, Sweden). Additional arterial blood samples (75 μL) were collected at 31, 32, 33, 34, 35, 36, 38, 42, and 46 min with respect to the glucose tracer bolus. Blood samples were centrifuged at 4°C, and two plasma aliquots were pipetted: 25 μL into 2 mL extraction mixture (for determination of 14C-2DG, 13C-glucose, 3H-R-BrP, and 14C-P levels) and 10 μL onto a cellulose cone for combustion (for determination of total 3H and 14C-levels, see below). After collection of the final blood sample rats were given an overdose of thiobutarbital, and tissues were rapidly dissected and frozen in liquid N2. Tissue samples were stored at −80°C awaiting analysis for tracer content (see below) and in the case of
