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. 2022 Feb 10;41:59. doi: 10.1186/s13046-022-02258-9

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Analysis of SMAD4 expression in HPV-positive HNC cell lines and tumours. A Total RNAs from HNC cell lines were isolated for RT-qPCR. SMAD4 expression was normalized to RpP0 and shown as means ± SD of at least three independent experiments. B Box plot showing means ± SD and individual values of results shown in (A). C Total RNAs from frozen tissue samples of 5 HPV-negative and 3 HPV-positive HNC patients were isolated for RT-qPCR. SMAD4 expression was normalized to RpP0 and shown as Box plot of the means ± SD and individual values. D HNC cell lines were lysed and analysed by immunoblotting with the indicated antibodies. E Box plot showing means ± SD and individual values of Optical density analysis of the expression of SMAD4 normalised to loading control (vinculin) from at least three independent western blot experiments in HNC cell lines. **, P < 0.01 (unpaired t test). F Box plot showing means ± SD and individual values of Optical density analysis from immunoblotting of SMAD4 expression from frozen tissue samples of 4 HPV-negative and 3 HPV-positive HNC patients