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. 2021 Feb 15;12:1042. doi: 10.1038/s41467-021-21356-4

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© The Author(s) 2021

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. 3 — a H&E-stained representative images in ileal sections of newborn mice, supplemented with AHR ligand I3C (25 mg per kg body weight per day for 4 days) and induced to develop experimental NEC. Data showing I3C mediated protection against NEC development only in wild-type (WT) and AHR myeloid knockout (Ahr^Δlys) mice but not in AHR knockout (Ahr^-/-) and AHR intestinal epithelial cells knockout (Ahr^ΔIEC) mice. b Dot graph showing AHR ligand I3C supplementation (25 mg per kg body weight per day for 4 days) produced a multifold induction of AHR activation marker Cyp1a1 in the ileum of wild-type but not in Ahr^-/-, and Ahr^ΔIEC and a moderate increase in Ahr^Δlys mice (n = 5, 11, 13, 11, 11 mice, Ctrl -I3C vs WT + I3C p < 0.0001, WT + I3C vs Ahr^Δlys + I3C p < 0.0001, Ahr^-/- + I3C vs Ahr^ΔIEC + I3C p = 0.0002, Ahr^ΔIEC + I3C vs Ahr^{Δlys +} I3C p = 0.0104). c–e NEC severity (c, n = 7, 7, 5, 7, 7 mice, Ctrl +I3C vs WT NEC + I3C p = 0.0020, WT NEC + I3C vs Ahr^-/- NEC + I3C p < 0.0001, WT NEC + I3C vs Ahr^ΔIEC NEC + I3C p < 0.0001, Ahr^ΔIEC NEC + I3C vs Ahr^Δlys NEC + I3C p = 0.0402) and mRNA levels of pro-inflammatory cytokine Il6 (d, n = 10, 16, 5, 10, 6 mice, WT NEC + I3C vs Ahr^-/- NEC + I3C < 0.0001, WT NEC + I3C vs Ahr^ΔIEC NEC + I3C p = 0.0001, Ahr^ΔIEC NEC + I3C vs Ahr^Δlys NEC + I3C p = 0.0086) and Tnf-α (e, n = 10, 16, 5, 10, 6 mice, WT NEC + I3C vs Ahr^-/- NEC + I3C p < 0.0001, WT NEC + I3C vs Ahr^ΔIEC NEC + I3C p < 0.0001, Ahr^ΔIEC NEC + I3C vs Ahr^Δlys NEC + I3C p = 0.0004) in the ileum of control mice without NEC and wild-type, Ahr^-/-, Ahr^ΔIEC, and Ahr^Δlys mice with NEC with I3C supplementation (25 mg per kg body weight per day for 4 days). Scale bars in a, 100 μm. All data are presented as mean values ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, p values obtained from two-sided t tests or one-way ANOVA followed by multiple comparisons. Each dot in graphs represents data from an individual mouse.