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. 2023 Jul 13;42:167. doi: 10.1186/s13046-023-02737-7

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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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — CD271 expression in cSCC biopsies and spheroids a Hematoxylin and Eosin (H&E) staining of AK, in situ, WD and MD/PD cSCC biopsies. CD271, Cyclin D1, TrkA, KRT1 and KRT13 expression evaluated by immunohistochemistry or immunofluorescence. Scale bar 100 µm. b Immunohistochemistry and immunofluorescence staining scored as follow: 0 (no cells; positivity to 20% of cells positivity), 0.1–0.75 (30–50% of cells positivity; 1–1.5 (60% to 100% of cells positivity). c Graphical representation of the KRT13/KRT1 intensity ratio. d Representative pictures of in situ, WD, and MD/PD spheroids. Scale bar = 100 μm. e Spheroid total area measured by ImageJ software and f spheroid viability evaluated by MTT assay. g Expression of E-cadherin, KRT13, and KRT10 evaluated in patient-derived spheroids by western blotting. β-actin was used as control. h Graphical representation of KRT13 and KRT10 relative protein expression ratio. i NTs and NTRs mRNA expression evaluated in patient-derived spheroids by qPCR. Heatmap created by Prism Graph pad software. β-actin was used as a housekeeping gene. For all experiments, the results are shown as mean ± SD of three independent experiments. Statistical analysis was performed using the two-way ANOVA. *0.01 < p < 0.05, ***0.0001 < p < 0.001, ****p < 0.0001