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. 2016 Jan 1;27(1):75–89. doi: 10.1091/mbc.E15-05-0319

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© 2016 Sun et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 10: — The insulin-stimulated colocalization of MC-Rab13 with both ACTN4 and GLUT4myc at cell periphery is reduced in cells expressing MICAL-L2-CT. (A) L6-GLUT4myc cells cotransfected with either GFP vector and MC-Rab13 or GFP-MICAL-L2-CT and MC-Rab13 were stimulated with insulin or not, fixed, labeled for ACTN4, and analyzed by spinning-disk confocal fluorescence microscopy. Top, collapsed optical z-stack images of GFP (green, inset), MC-Rab13 (red), and ACTN4 (blue). Bottom, images of GFP-MICAL-L2-CT (green, inset), MC-Rab13 (red), and ACTN4 (blue). Graphs show Pearson coefficients of colocalization of MC-Rab13 and ACTN4 at the cell periphery (mean ± SE, **p < 0.01). (B) L6-GLUT4myc cells treated as described were labeled for GLUT4myc and otherwise analyzed as before. Top, images of GFP (green, inset), MC-Rab13 (red), and GLUT4myc (blue). Bottom, images of GFP-MICAL-L2-CT (green, inset), MC-Rab13 (red), and GLUT4myc (blue). Graphs show Pearson coefficient of colocalization of MC-Rab13 and GLUT4myc at the cell periphery (mean ± SE, **p < 0.01, n = 3). Results are representative of three independent experiments (>25 cells/condition per experiment). Scale bars, 10 μm.