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. 2018 Apr 13;8:5949. doi: 10.1038/s41598-018-24503-y

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© The Author(s) 2018

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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Analysis of cell death mechanism after hTERT gene knockdown in HNSCC cell lines. (A) siRNA - Analysis of apoptosis- and autophagy-related genes expression at the transcriptional level with qPCR; (B) siRNA - Analysis of apoptosis- and autophagy-related genes expression at the protein level by Western blot analysis. Semi-quantitative analysis of Western blot results using ImageJ software; (C) Apoptosis analysis by flow cytometry; (D) shRNA - Analysis of apoptosis- and autophagy-related genes expression at the transcriptional level with qPCR; (E) shRNA - Analysis of apoptosis- and autophagy-related genes expression at the protein level by immunofluorescence staining, objective magnification 20x. CTRL – control cells transfected with non-specific siRNA/cells transduced with control lentiviral vector; siTERT – cells transfected with hTERT gene-targeted siRNA on day 3^rd and 7^th; shTERT – cells transduced with lentiviral vectors containing shRNA on day 10^th. Data are presented as the mean ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 with comparisons indicated by lines.