Abstract
Intrinsic to measuring insulin-stimulated GLUT4 translocation to the plasma membrane (PM) using total internal reflection fluorescence microscope (TIRFM) is the inability to differentiate intensity increment contributed by vesicles approaching the PM and GLUT4 molecules being inserted into the PM. Here, we combined IRAP-GFP probe, which exposes GFP to extracellular environment after GLUT4 vesicles fuse with the PM, with bromophenol blue (BPB), which quenches GFP fluorescence when in contact with it, trying to delineate the contribution of these two steps. It is found that insulin stimulation dramatically increased IRAP molecules localizing on the PM whereas GLUT4 vesicle density beneath the PM had little change through the translocation process. Therefore, we suggest that the fusion efficiency of GLUT4 vesicles at the PM is enhanced so much by insulin that GLUT4 vesicles barely accumulate underneath the PM.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
