Extended Data 7. Role of AHR in astrocytes and microglia during EAE.
(a-c) EAE was induced in Control (WT), GFAP-Cr AHRflox/flox (GFAP-AHR), or CX3CR1-AHR EAE mice. Starting from day 7, mice were injected daily intraperitoneally with Indoxyl-3-sulfate (I3S), given a tryptophan depleted diet (TDD), or kept on a control diet (Control). Clinical course of EAE mice under treatment conditions as indicated. Representative of 2 independent experiments with n = 4 mice per group. Data are mean ± s.e.m. and P values were determined by two-way ANOVA. (d-f) EAE was induced in WT mice, which were treated with lentiviruses to knock-down AHR in astrocytes (pGFAP-shAhr) or microglia (pCD11b-shAhr). A noncoding RNA was used as control. Flow cytometry quantification of AHR expression in astrocytes and microglia by FACS. (d) Representative histograms of n=4 mice per group. Numbers indicate percentage of AHR positive cells, thin lines isotype control, thick lines AHR staining. (e) Quantification of AHR positive astrocytes and microglia as in (d). Data are mean ± s.e.m. and P values were determined by one way ANOVA followed by Tukey’s post-hoc test. Representative of 2 independent experiments with n = 4 biological replicates. (f) EAE mice with knock down of AHR in astrocytes, microglia, or both as in (d) were subjected to daily I3S injections, TDD, or Control diet conditions starting on day 14 after disease induction. Clinical course of n=4 mice per group. Representative of 2 independent experiments with n = 4 mice per group. Data are mean ± s.e.m. and P values were determined by two-way ANOVA. (g) Quantification of CNS infiltrating pro-inflammatory monocytes as determined by FACS at day 28 of EAE. Data are mean ± s.e.m. and P values were determined by one way ANOVA followed by Tukey’s post-hoc test. Representative of 2 independent experiments with n = 3 biological replicates.