Abstract
Phospholipid fatty acid (FA) composition influences the biophysical properties of the plasma membrane and plays an important role in cellular signaling. Our previous work has demonstrated that plasma membrane fatty acid composition is an important determinant of oncogenic Ras signaling and that dietary (exogenous) modulation of membrane composition may underlie the chemoprotective benefits of long chain n-3 polyunsaturated fatty acids (PUFA). In this chapter, we describe in vitro methods to modulate membrane phospholipid fatty acid composition of cultured cells using fatty acids complexed to bovine serum albumin (BSA). Furthermore, we describe a method to quantify the biophysical properties of plasma membranes in live cells using Di-4-ANEPPDHQ (Di4) and image-based flow cytometry.
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Acknowledgments
This work was supported by the Allen Endowed Chair in Nutrition and Chronic Disease Prevention and the National Institutes of Health (R35-CA197707 and P30-ES029067). Natividad R. Fuentes is supported by the Texas A&M University Regulatory Science in Environmental Health and Toxicology Training Grant (T32-ES026568) and former recipient of a Predoctoral Fellowship in Pharmacology/Toxicology from the PhRMA Foundation. Michael L. Salinas is a recipient of the National Science Foundation Texas A&M University System Louis Stokes Alliance for Minority Participation (TAMUS LSAMP) Bridge to the Doctorate Fellowship (HRD-1612776).
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Fuentes, N.R., Salinas, M.L., Wang, X., Fan, YY., Chapkin, R.S. (2021). Assessment of Plasma Membrane Fatty Acid Composition and Fluidity Using Imaging Flow Cytometry. In: Rubio, I., Prior, I. (eds) Ras Activity and Signaling. Methods in Molecular Biology, vol 2262. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1190-6_14
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DOI: https://doi.org/10.1007/978-1-0716-1190-6_14
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