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. 2021 Feb 18;11:4181. doi: 10.1038/s41598-021-83418-3

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© The Author(s) 2021

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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The proportion of apoptotic cancer cells increases in PTCs showing ciliary loss. (A) Upon histological examination after H & E staining, apoptotic cancer cells appeared dark, with an eosinophilic cytoplasm and dense purple nuclear chromatin fragments (insert). TUNEL analysis revealed significantly more apoptotic cancer cells in PTCov and PTC-HT tissues than in PTC-conv tissue (d, e, and f). BCL-2 expression levels were lower in PTCov and PTC-HT tissues than in PTC-conv tissue (g, h, and i). Immunoexpression of VDAC1 was higher in PTCov and PTC-HT than in PTC-conv (j, k, and l). Scale bar, 10 μm. (B, C) Immunofluorescence images show labeling of primary cilia in normal human thyroid follicles and PTCs by antibodies specific for GT335 and ARL13B. Fewer primary cilia were detected in oncocytic cancer cells in PTCov and PTC-HT tissue than in normal thyroid follicles or conventional PTC tissue. Scale bar, 10 μm. Abbreviations: PTC-conv, conventional papillary thyroid carcinoma; PTCov, oncocytic variant of PTC; PTC-HT, PTC with Hashimoto’s thyroiditis background. ***P < 0.001, NS; not significant. (D) TUNEL with ARL13B double immunofluorescence staining revealed that TUNEL-positive cancer cells (green) have no primary cilia (stained by ARL13B; red).