TG synthesis was measured as described before18 (link). Briefly, synthesis of triglycerides derived from [14C]-oleate (PerkinElmer) complexed with BSA was measured by incubation of untreated or OL-treated cells with [14C]-oleate-BSA (0.2 μCi, 4 h), followed by extraction of cellular lipids and analysis by thin-layer chromatography. Incorporation of radioactivity into TG was determined using a Storm Gel and Blot Imaging System (GE Healthcare) followed by densitometry using ImageJ. The rate of TG synthesis was calculated as the amount of incorporated [14C]-oleate in relative units per mg protein.
Protocol detail
Measuring Triglyceride Synthesis in Cells
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Cellular
Densitometry
Lipids
Oleate
Protein
Radioactivity
Synthesis
Thin layer chromatography
Triglycerides
Corresponding Organization :
Other organizations : Albert Einstein College of Medicine
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Variable analysis
independent variables
- Treatment with OL
- Triglyceride (TG) synthesis rate
- Incorporation of [14C]-oleate into TG
- Incubation of untreated cells with [14C]-oleate-BSA
- Incubation time (4 h)
- Positive control: Untreated cells incubated with [14C]-oleate-BSA
- Negative control: Not explicitly mentioned
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