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. 2020 Jun 16;10:9705. doi: 10.1038/s41598-020-66257-6

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© The Author(s) 2020

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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(A) Representative FIHC images of control (non-progressing) and progressing hyperkeratosis and HGD showing DAPI channel on the left and merged PD-1(green)/PD-L1(red) on the right. The blue mask represents an automated cell identification protocol by Definiens Tissue Studio 4.0 based in DAPI staining. (B) Quantification of cell density per field, ANOVA and Tukey’s test, n = 10 fields per sample, ***P < 0.001.