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Insulin-induced cortical actin remodeling promotes GLUT4 insertion at muscle cell membrane ruffles
Peter Tong, Zayna A. Khayat, Carol Huang, Nish Patel, Atsunori Ueyama, Amira Klip
Published in Volume 108, Issue 3
J Clin Invest. 2001; 108(3):371–381 doi:10.1172/JCI12348
This article Copyright © 2001, The American Society for Clinical Investigation
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Figure 1

Confocal scanning series of GLUT4 colocalizing with newly formed dorsal actin-rich structures in response to insulin. Serum-deprived L6 GLUT4myc myotubes were treated with or without 100 nM insulin for 10 minutes at 37°C. Afterward, the cells were fixed, permeabilized, and double-stained for actin (Oregon Green-phalloidin) and GLUT4myc (anti-myc antibody followed by Cy3-conjugated secondary antibody) as described in Methods. Serial optical sections of 0.67 μm thickness along the z-axis were generated from myotubes as described in Methods. Optical slices taken from the dorsal to the ventral cell surface of myotubes show filamentous actin (a) and myc (b) after 10 minutes of 100 nM insulin treatment. The multiple myonuclei (b, dark ovals) were identified in parallel experiments by DAPI staining (data not shown). The progression of images is linear from left to right. Scale: 5 mm in image is equivalent to 10 μm in specimen. White arrows indicate regions of remodeled actin filaments. The images are representative of five experiments.

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