Supplementary Figure 3: AHR expression in WT and AHRLysM mice.

(a) Ahr expression in sorted bone marrow inflammatory monocytes (CD3^NegB220^NegLy6G^Neg NK1.1^NegSiglec-F^NegCD11b⁺Ly6C^Hi) from naive WT and AHR^LysM mice (n = 3 independent mice). Representative of two independent experiments with similar results. (b) Western-blot analysis of AHR in BMDMs from WT and AHR^LysM mice (n = 3 biologically independent samples). Data are representative of two independent experiments with similar results; images were cropped and the full scans are shown in Supplementary Fig. 2b. (c-e) qPCR analysis of Ahr expression in microglia (c), B cells (CD19⁺) and T cells (CD3⁺ CD4⁺ and CD3⁺ CD8⁺) (d), neutrophils (CD115^NegCD11b⁺CD11c^NegLy6G⁺) and dendritic cells (CD115^NegCD11b^NegCD11c⁺ MHCII^Hi) (e) (n = 3 independent mice). (f) Frequency of neutrophils and dendritic cells in the CD45⁺CD11b⁺ gate of glioma-infiltrating cells. CyTOF analysis was performed in WT mice 15 days after GL261 cell implantation. The dot plot graphs are representative of two independent experiments with similar results. (g) Frequency of microglia, splenic B cells and T cells in naive WT and AHR^LysM mice (n = 4 independent mice). (h) Total number of macrophages (CD11b+F4/80⁺) and inflammatory monocytes (CD11b⁺F4/80⁺Ly6C^Hi) in spleen from WT and AHR^LysM (n = 4 independent mice). (i) Survival curve analysis from WT and AHR^LysM mice implanted intracranially with CT2A cells (n = 10 independent mice). Representative of two independent experiments with similar results. Survival analysis was performed using a Kaplan-Meier plot using a log-rank (Mantel-Cox) test. (j) Flow cytometry analysis of TMEM119 (blue), CX3CR1 (red) and Ly6C (magenta) expression in TAMs, 15 days after GL261 implantation in WT mice. Representative of two independent experiments with similar results. (k) Ly6C expression on TAMs gate (Lin^NegCD11b⁺CD45⁺) in GBM from WT and AHR^LysM mice on day 15 (n = 3 independent mice). In the left, representative dot plot graphs of Ly6C expression in TAMs gate, where microglia (CD45^LowCD11b⁺) is shown in blue and peripheral infiltrated macrophages (CD45^HiCD11b⁺) in red. Percentage of Ly6C⁺ cells in TAMs (right panel). Representative of two independent experiments with similar results. (l, m) Ccl2 and Ccl7 gene expression in naïve brain and GL261 tumor-bearing mice (l) and in CT2A cells (m) (n = 3 tmice per group). Representative of two independent experiments with similar results. (n) Quantification of CCL2 in TCM from human GBM cells by ELISA (n = 5 biologically independent samples). Unpaired two-tailed t test was used to compare two groups (a-d, g, h, k, m and n) and one-way ANOVA was used to compare three or more groups (e and l). All data are presented as mean ± s.e.m.