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Updated: Sep 29, 2025

In vivo and in vitro Studies of Adaptor-clathrin Interaction
Published on: January 26, 2011
Evaluating the Role of Galectins in Clathrin-Independent Endocytosis
Mohit P Mathew1, Julie G Donaldson2, John A Hanover3
1National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, USA. mohit.mathew@nih.gov.
Abstract:
Galectin-3 is a chimeric galectin involved in diverse intracellular and extracellular functions. Galectin-3 is synthesized in the cytoplasm and then released extracellularly by a poorly understood non-canonical secretion mechanism. As a result, it can play important roles both inside and outside the cell. One important extracellular role of galectin-3 is in modulating clathrin-independent endocytosis (CIE), a form of cellular internalization that is still not well understood. CIE, unlike clathrin-mediated endocytosis, has neither defined signaling sequences nor cytoplasmic machinery. As a result, extracellular interactions like the galectin-glycan interactions are thought to directly drive changes in CIE. This chapter discusses the methods designed to study the role of galectin-glycan interactions in CIE, which have provided us with insight into the functions of galectin-3 and cell surface glycans during CIE cargo internalization. These methods include media supplementation for metabolic glycoengineering, antibody internalization assays, lectin panels to assay changes in glycan patterns, exogenous galectin-3 supplementation, galectin-3 secretion assays, and in vitro assays to monitor the effect of galectins on CIE.
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