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