Fig 1: a7nAChR is required for NNK-induced IDO1 upregulation and Trp metabolism. a The expression of the nine NNK receptors in 16HBE, A549, EPLC-32M and LLC cells. b The expression of a5nAChR- and a7nAChR-coding genes in A549 cells treated with 25 µM NNK for 72 h. c A549 cells were transfected with or without siCHRNA5 or siCHRNA7 and treated with 25 µM NNK for 72 h, and the expression of IDO1 was detected by qRT-PCR. d, e The cells were pretransfected with siCHRNA7 (d) or pretreated with a-BGT (e), followed by treatment with 25 µM NNK for 72 h. The cells were then lysed and subjected to western blotting using the indicated antibodies. f The A/J mice were treated with NNK and/or a-BGT for 90 days and sacrificed, and lung tissues were collected for western blot analysis. g The cells were transfected with siIDO1 and treated with 25 µM NNK for 72 h, and the Trp/Kyn ratio was tested. h The cells were transfected with siCHRNA7 and treated with 25 µM NNK or 10% CSE for 72 h, and the Trp/Kyn ratio was tested. P values, Student’s t test. Error bars, sd
Fig 2: Fludarabine Antagonizes IFN-?-Induced STAT3 Activation and Upregulation of IDO1 in HCC Cells(A) LM3 cells were treated with IFN-? (2,000 IU/mL) for 2, 12, or 24 h, lysed, and STAT3/p-STAT3 protein levels were determined by western blotting. (B and C) LM3 cells were treated with fludarabine (200 nM) and IFN-? (2,000 IU/mL) for 12 (B) or 24 (C) h in the presence of MG132 (10 µM) and lysed, and STAT3 activation (B) and IDO1 expression (C) were assayed by western blotting. GAPDH was used as a loading control. Representative blots of two independent experiments are shown.
Fig 3: miR-155 upregulates TNF-a, TNF-ß and IDO1 expression in BV-2 microglial cells. BV-2 microglial cells were transfected with mimic control, miR-155 mimics or treated with LPS. Western blotting was performed to evaluate the expression levels of TNF-a, TNF-ß and IDO1 in BV-2 microglial cells. #P<0.05 vs. control; **P<0.01, ***P<0.001 vs. mimic control. miR, microRNA; LPS, lipopolysaccharide; TNF, tumor necrosis factor; IDO1, indoleamine 2,3-dioxygenase 1.
Fig 4: IFN-? induces IDO activity in the HeLa cell lines, the activity of which is inhibited by different IDO inhibitors (IDO49, IDO5l, IDO5m)
Fig 5: Expression IDO1 of the miceMice injected with IFN-? (7.5 µg/day on days 1, 2, 3, 4, 5, 8) and tumor tissues were collected. Protein was extracted from the tumor for the western blotting analysis. (“+” represent inject IFN- ?, “-” represent non-inject IFN-?).
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