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. 2023 Jan 9;14:116. doi: 10.1038/s41467-022-35741-0

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© The Author(s) 2023, corrected publication 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — A Color-encoded Z-stack of wt and Pg-S665A keratinocytes revealed multiple layers in wt and only 1-2 layers in Pg-S665A keratinocytes. Representative of n = 3. Scale bar = 50 µm. A, B Dsg1 was expressed primarily in the superficial epidermal layers in a linear fashion along cell borders. In contrast, Dsg1 was drastically reduced and restricted to the basal layer in Pg-S665A keratinocytes. C Pg was reduced and appeared less linearized in Pg-S665A keratinocytes compared to wt. D pPg-S665 was absent in Pg-S665A keratinocytes confirming phospho-deficiency at this site whereas it appeared dotted along cell borders and in the nuclei in wt keratinocytes. E The keratin cytoskeleton showed a dense network spanning the whole cell in wt keratinocytes whereas it was rarefied and drastically altered in Pg-S665A keratinocytes. B–E Scale bar = 25 µm. White rectangles depict areas for zoom in. Scale bar (zoom) = 5 µm. Representatives of n = 4 (B), n = 3 (C–E). F Triton-based separation revealed a drastic impairment of Dsg1 and Dsg3 expression in the Triton insoluble, desmosome bearing fraction in Pg-S665A keratinocytes. Similar observations were made for Dp and Pg. Importantly, keratin 14 (CK14) was almost absent in Pg-S665A keratinocytes. In contrast, β-catenin was slightly upregulated in the Triton insoluble fraction. Tubulin serves as loading control. Representative of n = 4. G Dissociation assay in wt and Pg-S665A keratinocyte cell lines showed a drastic impairment of intercellular adhesion in Pg-S665A cells (wt-1, Pg-S665A-2: n = 6; wt-2, Pg-S665A-1: n = 4). H Dissociation assay in wt murine keratinocytes revealed a significant fragmentation after 24 h of incubation with the pathogenic monoclonal anti-Dsg3 antibody AK23. Pre-incubation with apremilast (Apr) for 1 h ameliorated AK23-induced loss of intercellular adhesion in wt keratinocytes (n = 4, for vehicle: n = 3). In contrast, in Pg-S665A keratinocytes (I), which already showed drastic impairment of intercellular adhesion under resting conditions, AK23 had no additional effect on intercellular adhesion (n = 7). Apremilast did not restore impaired intercellular adhesion neither under basal conditions nor after AK23 incubation. Columns indicate mean value ±SEM. P < 0.05 (exact values are depicted in the figure). One-way ANOVA with Bonferroni correction. Source data are provided as a Source Data file.