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. 2001 Jun 25;153(7):1369–1380. doi: 10.1083/jcb.153.7.1369

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Measurements of Akt-PH–GFP FRAP. RAW 264.7 cells transfected with Akt-PH–GFP were resuspended and allowed to sediment on IgG-coated coverslips as in the legend to Fig. 5. Confocal images of the adherent membrane were acquired before (A, Pre), immediately after (B, 0 s), and at the indicated intervals following photobleaching of a spot near the middle of the membrane (C–D). In E, the fluorescence intensity along line scans traversing the photobleached area was determined at the indicated times after photobleaching. The width of the initial bleached area is demarcated by the dotted lines. In F, the relative fluorescence intensity at the center of the area selected for analysis is plotted versus time after bleaching. Data are representative of five experiments.