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. 2018 May 17;9:1981. doi: 10.1038/s41467-018-04366-7

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© The Author(s) 2018

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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Fig. 5 — System L-mediated transport of kynurenine drives AHR signalling. a CD4⁺ T cells were activated and maintained in TH17 conditions (CD3/CD28; IL-6, IL-1β and TGFb) (TH17^cond). Uptake of ³H-kynurenine in the presence or absence of BCH (5 mM) or non-radiolabelled kynurenine (5 mM) at times D2, D3 and D5 post activation is shown. b Histograms showing 450 nm fluorescence of D0, D3 and D5 TH17^cond CD4⁺ T cells treated with kynurenine (200 μM, 4 min) in the presence or absence of BCH (5 mM), or with no kynurenine. c Ratio of the 450 nm emission of D0, D3 and D5 TH17^cond CD4⁺ T cells treated with kynurenine ± BCH 5 mM (4 mins, 37 °C). d The graph shows the mean fluorescence intensity (MFI) of D3 TH17^cond CD4⁺ T cells treated with kynurenine (100 μM, 4 min, emission 450 nm) or FICZ (300 nM, 4 min, emission 530 nm) in the presence or absence of BCH (10 mM). e, f Representative flow cytometry histograms of (d); kynurenine uptake (e) and FICZ uptake (f). g The graph shows Cyp1a1 mRNA from D3 or D5 TH17^cond CD4⁺ T cells stimulated with kynurenine (100 μM, 2 h) or FICZ (300 nM, 2 hr), relative to non-treated control. h Induction of Cyp1a1 mRNA in D3 TH17^cond CD4⁺ T cells stimulated with kynurenine (100 μM, 2 h) or FICZ (300 nM, 2 h) in the presence or absence of BCH (25 mM). The data are shown relative to non-treated control. i Dose–response induction of Cyp1a1 mRNA in D3 TH17^cond CD4⁺ T cells stimulated with kynurenine at indicated doses for 2 h. The data are shown relative to non-treated control. j Induction of Cyp1a1 mRNA in D3 TH17^cond CD4⁺ T cells stimulated with kynurenine (10 μM, 2 h) in the presence or absence of AHR antagonist (CH223191, 30 μM). The data are shown relative to non-treated control. Error bars are ±s.d. Individual points indicate biological replicates. MFIs are indicated in the histograms, data are from three biological replicates (a–g, i–j) and six (Kyn) or three biological replicates (FICZ) (h). P values * = < 0.01; ** = < 0.005; *** = < 0.001; **** = < 0.0001; ns= not significant (ordinary one-way ANOVA)