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. 2015 Jul 1;26(13):2402–2417. doi: 10.1091/mbc.E15-01-0043

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© 2015 Ketema 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 6: — HD composition, cytoskeletal organization, and integrin β4 phosphorylation are normal in rodless plectin keratinocytes. (A) Wild-type (clone 12) and rodless (clone 16) plectin keratinocytes were grown on collagen I–coated coverslips. Cells were fixed in paraformaldehyde and stained for integrin β4 and either plectin (top) or BP230 (bottom). Nuclei were counterstained with DAPI, and cells were analyzed by confocal microscopy. Scale bars, 20 μm. (B) The cell surface expression of integrin β4 (top) and integrin α6 (bottom) was investigated by flow cytometry in wild-type (clone 12) and rodless (clone 16) plectin keratinocytes. Gray shading indicates cells incubated with only secondary antibody. (C) Time course of EGF-induced phosphorylation of integrin β4 at S1356 and S1364 in wild-type (clone 12) and rodless (clone 16) plectin keratinocytes. Cells were deprived of growth factors for 24 h and subsequently stimulated with EGF for the indicated times. Cell lysates were analyzed by Western blot for expression of the indicated proteins. (D) Wild-type (clone 12) and rodless (clone 16) plectin keratinocytes were treated as described in A and stained for plectin, integrin β4, and either F-actin (top) or keratin 14 (bottom). Cells were subsequently analyzed by confocal microscopy. Scale bars, 20 μm.