Clathrin-mediated endocytosis (CME) is important for organismal growth, nutrient uptake and pathogen defence, but the molecular characteristics of CME in plants has remained largely unexplored. To this end and to support the systematic discovery of CME function in planta, in their Tools and Resources article, Jiří Friml and colleagues (Johnson et al., 2020) now provide an experimental toolbox for the quantitative evaluation of CME in Arabidopsis. Among the standardized protocols, they include a detailed guide to the imaging of Arabidopsis CME vesicles through electron microscopy in order to characterize their ultrastructure. In addition, the authors also evaluate the major light microscopy methods used for CME studies; this is accompanied by a detailed protocol to quantitatively analyse the temporal events during endocytosis and the interplay of components in single CME events. Furthermore, they describe a method for the quantitative characterisation of the global internalisation of cargoes; this employs a semi-automated MATLAB analysis tool and can be used with FM dyes to visualise membrane internalisation events during endocytosis. Finally, the authors present an overview of available pharmacological and genetic tools for CME studies on intact plants, together with some validation. Overall, this toolbox will not only be instrumental in extending research of CME in plants, but also provide an easy to use and go-to guide for researchers embarking on studying CME for the first time.
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