Abstract
In eukaryotic cells, nuclear envelope transmembrane proteins (NETs) are synthesized on the endoplasmic reticulum and then transported from the outer nuclear membrane (ONM) to the inner nuclear membrane (INM). Quantitative determination of the distribution of NETs on the ONM and INM is limited in available approaches, which moreover provide no information about translocation rates in the two membranes. Furthermore, commonly used techniques, such as electron microscopy, are time-consuming, and performed in vitro. Here, we demonstrate a single-point FRAP microscopy technique that enables determination of distribution and translocation rates in vivo with <10-nm spatial resolution within 30 minutes.
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