Abstract
Clathrin-coated pits are major ports of entry into eukaryotic cells. Following scission of clathrin-coated pits to form coated vesicles, the peripheral membrane proteins that form the coat need to be removed. Here we describe an assay that provides a measure of the extent of clathrin coat uncoating in intact cells. This assay has been used to investigate rab5-dependent modulation of uncoating. Specifically, it has been used to identify functional differences between rab5 guanine nucleotide exchange factors.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Biological Assay / methods*
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Cell Culture Techniques
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Clathrin-Coated Vesicles / metabolism*
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Fluorescent Antibody Technique
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Gene Knockdown Techniques
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Guanine Nucleotide Exchange Factors / deficiency
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Guanine Nucleotide Exchange Factors / genetics
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Guanine Nucleotide Exchange Factors / metabolism*
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HEK293 Cells
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Humans
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Image Processing, Computer-Assisted
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RNA, Small Interfering / genetics
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Transfection
Substances
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GAPVD1 protein, human
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Guanine Nucleotide Exchange Factors
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RNA, Small Interfering