Tissue-specific fatty acid oxidation was measured by the production of 14CO2 from [1-14C]oleic acid (0.3 μCi/mL), with unlabeled oleate present in the medium as previously reported with minor modification (10 (link)). Mice were fed HFD for 6 weeks and fasted overnight. Soleus, extensor digitorum longus, and tibialis anterior muscles and liver tissue were weighed and placed in 50-mL glass flasks. The flasks had an isolated center well containing 1 N NaOH to trap 14CO2. After 30 min of incubation at 37°C, the medium was acidified with 1 mL of 0.5 N sulfuric acid to stop the reaction. Flasks were then held at 50°C for an additional 3 h. The 14C activity of the contents in the center well was counted in scintillation cocktail (Ultima Gold; PerkinElmer) by using a β-counter (Beckman Scintillation Counter; PerkinElmer).
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Tissue-Specific Fatty Acid Oxidation
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Fatty acid
Gold
Held
Liver
Mice
Oleate
Oleic acid
Scintillation counter
Soleus
Sulfuric acid
Tibialis anterior muscles
Tissue
Tissue specific
Corresponding Organization : Veterans Health Administration
Other organizations : Gachon University, University of Washington, Howard Hughes Medical Institute
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Variable analysis
independent variables
- Diet (HFD vs. control)
- Tissue-specific fatty acid oxidation (measured by production of ^14CO2 from [1-^14C]oleic acid)
- Unlabeled oleate present in the medium
- Incubation time (30 min at 37°C, followed by 3 h at 50°C)
- Tissue samples (soleus, extensor digitorum longus, tibialis anterior muscles, and liver)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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