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Leucine oxidation

RS Rasmus J. O. Sjögren
DR David Rizo-Roca
AC Alexander V. Chibalin
EC Elin Chorell
RF Regula Furrer
SK Shintaro Katayama
JH Jun Harada
HK Håkan K. R. Karlsson
CH Christoph Handschin
TM Thomas Moritz
AK Anna Krook
EN Erik Näslund
JZ Juleen R. Zierath
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Myotube cultures were incubated with 1 ml of Ham’s F10 Medium in the presence of 0.1 mCi/ml radiolabelled leucine (l-[U-14C]leucine; 11.1 GBq/mmol, NEC279E250UC; PerkinElmer, MA, USA) and either 3,6-dichlorobenzo(b)thiophene-2-carboxylic acid (BT2) (ENA018104907; Sigma-Aldrich) or DMSO. Small cups were placed in cell-culture wells and plates were air-tight sealed. After incubation for 4 h at 37°C, 150 μl of 2 mol/l HCl and 300 μl of 2 mol/l NaOH were added into each well and small cup, respectively. After 1 h, the liberated CO2 was collected and subjected to scintillation counting (Tri-Carb 4910TR; PerkinElmer).

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