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. 2014 Jan 20;5(4):1038–1051. doi: 10.18632/oncotarget.1637

Figure 2. STAT3 mediates constitutive IDO1 expression in vitro.

Figure 2

A, Cartoon of the human IDO1 promoter showing two STAT3 binding sites (pink) and two interferon-sensitive response elements (ISRE, green) upstream of the transcription start site. B, Western blot of STAT3 phosphorylation in indicated cell lines. STAT3 served as loading control. C, IDO1 mRNA, IDO protein and kynurenine release of SKOV-3 cells after 24 h (mRNA) or 48 h (protein and kynurenine) of treatment with 100 μM AG490. D, IDO protein and kynurenine realease of SKOV-3 cells analyzed after 24 h treatment with 1 μM JSI-124. E, IDO1 mRNA and IDO protein 72 h after STAT3 shRNA in SKOV-3 cells in comparison to control. F, Immunoprecipitation of STAT3 of indicated cell lines. Acetylation of lysine residue 685 of STAT3 was analyzed using western blot. STAT3 served as loading control. G, IDO1 mRNA, IDO protein and kynurenine release of SKOV-3 cells, analyzed 24 h (mRNA) or 48 h (western blot and HPLC) after treatment with 20 μM HATI. H, Mixed leukocyte reactions (MLR) on top of 2000 SKOV-3 cells, which were pre-treated with 100 μM AG490, 1 μM JSI-124 or 20 μM HATI for 2 days before addition of the MLR in comparison to controls. Asterisk indicates p<0.05, error bars indicate s.e.m.