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. 2021 Aug 9;19:338. doi: 10.1186/s12967-021-02987-z

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© The Author(s) 2021

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. 5 — CYB561D2-activated STAT3 induces the expression of immunosuppressive genes. A Representative western blots showing the protein levels of FASLG, TGF-β2, CD70, PD-L1, PD-L2, CCL2 and TDO2 after transfection with CYB561D2 plasmid at indicated dose (0, 0.5, 1, 2 μg/well in 6-well plate) in U251 and U87 cell lines. Inhibitor indicates STAT3 inhibitor C188-9. B qRT-PCR results showing the relative mRNA levels of PD-L1, CCL2 and TDO2 after transfection with CYB561D2 plasmid at indicated dose in U251 and U87 cell lines. C Scatter plots of gene expression showing the association of CYB561D2 with PD-L1 (Spearman r = 0.19, P = 0.0003), CCL2 (Spearman r = 0.33, P < 0.0001) and TDO2 (Spearman r = 0.29, P < 0.0001) in TCGA GMB dataset. D RealTime-Glo™ Annexin V Apoptosis assay showing the effect of CYB561D2 over-expressing U251 cells on co-cultured mouse T cells. E Caspase-Glo® 3/7 assay showing the effect of CYB561D2 over-expressing U251 cells on co-cultured mouse T cells. F IL-2 ELISA showing the effect of CYB561D2 over-expressing U251 cells on co-cultured mouse T cells. For all, *P < 0.05; **P < 0.01; ***P < 0.001