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. 2017 Aug 24;7:9373. doi: 10.1038/s41598-017-09668-2

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© The Author(s) 2017

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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Combination of NCTD with CLSO downregulates peripheral blood CD4 ⁺ CD25 ⁺ FoxP3 ⁺ Tregs in mice. (a) Isolated spleen weight from the Hepal-1 hepatoma-bearing model. (b) The peripheral blood cells were harvested, and the percentage of CD4 ⁺ CD25 ⁺ FoxP3 ⁺ cells was determined using flow cytometry. The data from the peripheral blood of mice are shown. (c) Gene expression analysis of FoxP3 and CTLA-4 of the peripheral blood of Hepal-1 mice by Quantitative Real-time PCR. The relative quantification value, fold difference, is expressed as 2^−ΔΔCt. (d) The expression levels of FoxP3 in the peripheral blood samples were detected by Western blot analysis. Results represent mean values of three experiments ± SD. (e) The concentrations of TGF-β and IL-10 in the serum of mice were measured by ELISA. Data are representative of three independent experiments.^*P < 0.05; ^**P < 0.01, vs. Vehicle, ^aP < 0.05; ^aaP < 0.01 vs. NCTD alone, ^bP < 0.05; ^bbP < 0.01 vs. CLSO alone. One-way ANOVA, post hoc comparisons, Tukey’s test. Columns, means; error bars, SDs.