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. 2023 May 12;13:7743. doi: 10.1038/s41598-023-33485-5

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© The Author(s) 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Increased resistance does not depend on alterations in desmosomes. (a) Dispase assay with stratified keratinocyte multilayers (48 h Ca²⁺) transfected with either ctr siRNA (siCtr) or siRNA against desmoglein3 (siDsg3). Quantification of sheet fragments upon end over end rotation: *P = 0.0116 with 1-way ANOVA, Tukey’s post test for Ctr/aPKCλ^−/−, siCtr/siDsg3: n = 5. (b) Immunofluorescence staining for β-catenin, desmoplakin (DP), desmoglein1/2 (Dsg1/2) and desmoglein3 (Dsg3) in stratified keratinocyte multilayers (48 h Ca²⁺). Enlargements show punctate desmosomal intercellular junctions. Nuclei labeled with DAPI (blue). (c) Quantification of sheet fragments upon EGTA treatment and end over end rotation. **P = 0.009 with 1-way ANOVA, Tukey´s post test for Ctr: n = 26, aPKCλ^−/−: n = 19, aPKC^dKO: n = 6. (d) Western Blot quantification of total and phospho-desmoplakin levels (Ser2849) in lysates of stratified keratinocyte sheets (48 h Ca²⁺). (e) Transmission electron micrographs, cross section of stratified keratinocyte sheets 48 h in high Ca²⁺ showing comparable desmosome in Ctr and aPKCλ^−/− keratinocytes. Representative images from n = 3 (Ctr/aPKCλ^−/−). (f, g) Quantification of desmosomal length (f) and quantity (g) from transmission electron micrographs as shown in (e) for Ctr/aPKCλ^−/−/aPKC^dKO: n = 3. Numbers of quantified desmosomes/biological replicate are indicated above the graph. (h) Dispase assay with stratified Ctr and E-cadherin^−/− keratinocyte multilayers (48 h Ca²⁺) after detachment upon dispase treatment. Graph: Quantification of sheet fragments upon sonication. n = 5 (Ctr), n = 6 (E-cadherin^−/−).