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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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Loss-of-function of primary cilia in thyroid cancer cell lines upregulates the mitochondria-dependent apoptosis pathway. (A) Expression of VDAC1, VDAC2, and VDAC3 mRNA in human thyroid cancer cells (TPC1 and BCPAP), with or without ciliary loss. **P < 0.01, ***P < 0.001, NS; not significant. (B) Comparison of immunofluorescence staining of VDAC1 between negative control siRNA-transfected cells and KIF3A-deficient TPC1 or BCPAP cells. Scale bar, 10 μm. (C) Western blot analysis of VDAC1 and HSP60 (a mitochondrial volume marker) expression in whole-cell lysates of KIF3A-KD or IFT88-KD TPC1 compared with that in negative control siRNA-transfected cells. (D) Analysis of the oligomeric status of VDAC1 in the mitochondrial fractions of KIF3A-KD and IFT88-KD TPC1 cells compared with negative control siRNA-transfected cells. A line graph was generated from the ImageJ data using arbitrary area units. *P < 0.05. (E) Western blot analysis of VDAC1 and HSP60 expression in whole-cell lysates of KIF3A-KD and IFT88-KD BCPAP cells compared with negative control siRNA-transfected cells. (F) Analysis of the oligomeric status of VDAC1 in the mitochondrial fraction of KIF3A-KD and IFT88-KD BCPAP cells compared with negative control siRNA-transfected cells. A line graph was generated from the ImageJ data using arbitrary area units. *P < 0.05, NS; not significant. (G, H) Flow cytometry analysis of apoptosis in IFT88-deficient TPC1/BCPAP cells treated with DIDS. The levels of apoptosis in IFT88-deficient TPC1 or BCPAP cells treated with DIDS were markedly less than those in untreated IFT88-KD cells. Bar graphs show average percentages of apoptotic cells (Q2 + Q4). **P < 0.01.