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. 2024 Feb 10;25:4. doi: 10.1186/s12860-024-00500-0

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© The Author(s) 2024

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/. 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. 6 — Expression of Shh and Nrf2 in blood and tissue samples based on CTCs. A Shh expression was quantified by qRT-PCR normalized against GAPDH in blood samples of HNSCC patients and compared with healthy control. B, C qRT-PCR was used to measure Shh expression in study subjects’ blood and tissue samples, normalized against GAPDH. The results were compared between patients who tested positive for CTC and those who did not. D, E qRT-PCR was used to quantify Shh expression in both CTC-positive and CTC-negative HNSCC patients. The results were normalized against GAPDH and compared between tissue and blood samples. F The expression of Nrf2 was measured in blood samples from study participants using qRT-PCR, normalized against GAPDH, and compared to healthy control subjects. G, H The expression of Nrf2 was also measured in study subjects’ blood and tissue samples using qRT-PCR, normalized against GAPDH, and compared between patients who tested positive for CTC and those who tested negative. I, J Quantitative real time PCR was carried out to determine Nrf2 expression in both blood and tissue samples from patients with HNSCC who were CTC-positive and negative, normalized against GAPDH. Data represented as mean ± SD and the Mann–Whitney test was carried out to compare Shh and Nrf2 expression among the study subjects. **p-value < 0.01, ****p-value < 0.0001, two-tailed unpaired t-test (Mann–Whitney test)