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. 2017 Sep 15;28(19):2492–2507. doi: 10.1091/mbc.E17-01-0022

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© 2017 Mathiowetz, Baple, 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 4: — Amish GMS fibroblasts have altered F-actin organization and ERGIC morphology. (A) Fibroblasts from two healthy (+/+) Amish individuals or Amish GMS (GMS/GMS) patients were stained with phalloidin to visualize F-actin (green) and with DAPI to label DNA (blue). Unusual orthogonal bundles of F-actin fibers and aggregates are highlighted with arrows. A lack of cytoplasmic F-actin puncta in GMS cells is exemplified in the row of magnified images. (B) The percentage of cells with aberrant (orthogonal, aggregated, or stellate) F-actin stress fibers was quantified. Each bar represents the mean (+ SEM) from analyses of 350 cells. ***, p < 0.001 (Fisher’s exact test). (C) The percentage of cells with prominent cytoplasmic F-actin puncta in the juxtanuclear region was quantified. Each bar represents the mean (+ SEM) from analyses of 350 cells. For cells with puncta, the number per cell was also quantified. Each bar represents the mean (+ SEM) from analyses of 12 cells. ***, p < 0.001 (Fisher’s exact test, ANOVA). (D) Wild-type or GMS fibroblasts were stained with antibodies to detect ERGIC-53 (red) and with DAPI to label DNA (blue). (E, F) The areas occupied by the ERGIC, nucleus, and entire cell were calculated in ImageJ and converted to a ratio of organelle area/cell area. Each bar represents the mean (+ SEM) from analyses of 25 cells. ***, p < 0.001 (ANOVA). All scale bars: 10 μm.