Dissecting GLUT4 Traffic Components in L6 Myocytes by Fluorescence-Based, Single-Cell Assays

Abstract

Postprandial blood glucose homeostasis is regulated by an insulin-stimulated increase in glucose transport into muscle and fat tissues via glucose transporter isoform 4 (GLUT4). In the basal state, this constitutively recycling membrane protein predominantly resides intracellularly. In order to achieve the insulin-stimulated increase in glucose flux, GLUT4 increases its cell surface abundance at the expense of preformed intracellular depots. By confocal microscopy of cultured L6 muscle cells stably expressing myc-tagged GLUT4 (L6-GLUT4myc), we can visualize the two arms of GLUT4 traffic: exocytosis (movement to the cell surface) and endocytosis (internalization from the cell surface).