Immunofluorescence Study of Endogenous YAP in Mammalian Cells
Immunocytochemistry enables determination of cellular localization and relative abundance of proteins. This protocol describes a rapid and cost-effective approach to study the cellular localization of YAP (and TAZ), the transcriptional co activators of the Hippo pathway, in mammalian cells. Cells are seeded onto coated cover slips, cultivated and treated as required. Subsequently, they are chemically fixed, and cellular proteins are fluorescently labeled by means of specific antibodies. Multiplexing antibodies enables ascertaining the subcellular localization of YAP and TAZ and thereby also the activation state of the Hippo pathway in various cell types.
Key wordsImmunofluorescence Immunocytochemistry Cellular localization YAP Hippo pathway
Members of the Gram Hansen lab are thanked for their comments as well as for helping taking the photos for Figs. 1, 2, 3, 4, 5, and 6. Work ongoing in the Gram Hansen lab is supported by a Chancellor’s Fellowship start-up fund and by the Wellcome Trust-University of Edinburgh Institutional Strategic Support Fund. We acknowledge the technical support and guidance provided by the Centre for Reproductive Health SuRF Histology and imaging as well as the Centre for Inflammation Research imaging staff.
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