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All living cells approach information by trafficking cargo, which include extracellular ligands, microorganisms, nutrients, transmembrane proteins and Cereblon Inhibitor custom synthesis lipids in the plasma membrane to endocytic vesicles (i.e. endocytosis). A reciprocal process known as recycling balances endocytosis and returns substantially from the internalized membrane and cargo for the cell surface. The balance between endocytosis and recycling controls the plasma membrane composition and gives cells with details that has been resolved in time and space. Endocytosis and recycling are master regulators of diverse cellular functions such as nutrient uptake and metabolism, improvement, proliferation, differentiation and polarity, 1-3 reprogramming, migration, cell adhesion and migration, cytokinesis, and neurotransmission . Endocytic and recycling pathways are very dynamic and highly coordinated and permit cells to turn over the equivalent with the complete plasma membrane 1-5x per hour. The cell-based L-glutahione protection assays are valuable to study endocytosis and recycling of transmembrane proteins like receptors, 4-8 channels, transporters, and adhesion molecules in epithelial and nonepithelial cells . We have previously studied endocytosis and recycling 9-15 in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) in human airway epithelial cells and HEK293 cells . The biotinylationbased assays described within the manuscript are optimized for examining endocytosis and recycling in epithelial cells cultured beneath polarizing conditions on semipermeable development supports. These protocols might be modified to study endocytosis and recycling of proteins in epithelial cells cultured in plastic tissue culture dishes or in nonepithelial cells. Figures 1 and two include examples of endocytic and recycling assays in epithelial and nonepithelial cells.