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. 2022 Nov 28;13:7148. doi: 10.1038/s41467-022-34815-3

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© The Author(s) 2022

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. 3 — A t-SNE depicting the global proteomic profile correlation between normal sinonasal tissue (CTRL), olfactory neuroblastoma (ONB), sinonasal squamous cell carcinoma (SCC), NEC-like IDH2, NEC-like SMARCA4/ARID1A, and adenoid cystic carcinoma (ACC). ONB, NEC-like IDH2 and NEC-like SMARCA4/ARID1A had an unexpectedly similar proteomic profile. B Differential expression analysis between normal sinonasal tissue and the investigated tumor classes was performed using a moderated t-test followed by Benjamini-Hochberg multiple testing correction. The results are shown as volcano plots. Recurrent and highly differential expressed proteins are annotated, highlighting overexpression of proteins specific for neurons or neuroendocrine cells in the ONB, NEC-like IDH2 and SMARCA4/ARID1A class. C Combined bar and point plots of 59 biologically independent samples showing the expression of Cytokeratin 18 (KRT18) and Ubiquitin carboxy-terminal hydrolase L1 (UCHL1) in the different tumor classes as determined by proteomics and immunohistochemistry (IHC). D Exemplary hematoxylin/eosin and KRT18 and UCHL1 immunohistochemical stainings are shown, validating the results from the proteomics analysis. In conclusion, the combination of these markers could be useful for histopathological classification if DNA methylation is not available or feasible. E Results from overrepresentation analysis comparing the overall similarity of the global protein expression signatures of tumor classes with various normal cell types of the airways. Differentially expressed genes between normal sinonasal tissue and the investigated tumor classes were subjected to overrepresentation analysis using previously published cell type-specific gene sets that were identified using single-cell RNA sequencing data of the mucosal lining of human airways. The Fisher’s exact test followed by FDR multiple testing correction was used to test for significance. A high similarity between pulmonary neuroendocrine cells (‘PNEC’) and the ONB, NEC-like IDH2 and NEC-like SMARCA4/ARID1A was observed, in line with the overexpression of neuronal/neuroendocrine markers shown in B. ACC specimens mostly resembled serous cells of submucosal glands and SCC specimens mostly resemble Squamous Cells 1. ACC adenoid cystic carcinoma, ADC adenocarcinoma, NEC neuroendocrine carcinoma, CTRL normal sinonasal tissue, ONB olfactory neuroblastoma, PDCA poorly differentiated carcinoma, SCC squamous cell carcinoma, FDR False discovery rate, PNEC Pulmonary neuroendocrine cells.