Abstract
Endocytosis impacts many cell biological functions, including in embryonic stem cells (ESCs). It has been shown that endocytosis is necessary for adequate FGF-signaling within the preimplantation ESC to post-implantation epiblast (EpiLC) pluripotency continuum and is required for proper levels of ERK activation. Quantitative methods at single cell resolution are needed to study endocytosis as well as its regulation and roles in these transitioning populations. The methods in this chapter provide an easily adaptable, multiplexable platform to monitor and quantify endosomal uptake at single cell resolution in live cells following receptor-mediated and non-receptor-mediated endocytosis, including nonspecific mechanisms such as pinocytosis.
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Acknowledgments
This work was supported by Eunice Kennedy Shriver National Institute of Child Health and Human Development grant support to C.S. (F32HD088051). Thanks to Robert Blelloch for support and critical analysis. Thanks to Kevin Chen and Kayla Lenshoek from the Blelloch laboratory for critical feedback.
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Sangokoya, C. (2022). Measuring Endocytosis and Endosomal Uptake at Single Cell Resolution. In: Osteil, P. (eds) Epiblast Stem Cells. Methods in Molecular Biology, vol 2490. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2281-0_6
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DOI: https://doi.org/10.1007/978-1-0716-2281-0_6
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