Fig. 3: VGLUT3 and EAAT2 are regulated with NRF2-AHR pathway in the ethanol-fed mice.

a Protein-protein interactions of L-glutamate transporter activity (GO:0005313) in bulk RNA-seq data of mouse livers through the STRING database. b Top 10 up-regulated genes stimulated by AHR agonists (TCDD, ITE) in mouse lung fibroblast, re-analyzed using public microarray data. c mRNA expression of NRF2 and AHR-related genes and AHR target genes from RNA-seq analysis of WT mice fed with Pair or EtOH diet for 2 weeks (n = 3/group). d Relative mRNA expression of AHR-related genes according to liver zonation in scRNA-seq of WT mouse liver (GSE84498). e Western blot of NRF2 and AHR in isolated HEPs. f Representative immunofluorescent staining of AHR and NRF2 in liver sections. Scale bar, 50 μm. g, h mRNA expression of Nfe2l2, Slc1a2, and Slc17a8 in freshly isolated HEPs treated with 20 mM EtOH and N-acetyl-L-cysteine (NAC) for 24 h (g, n = 4/group), along with their Western blot analysis of NRF2, VGLUT3, and EAAT2. i In situ closed liver perfusion performed with VEH, 100 mM EtOH, or 1 μM AHR agonist (ITE) containing media in WT mice (2 h). Relative mRNA (n = 6/group) and protein levels of Ahr (AHR), Nfe2l2 (NRF2), Slc17a8 (VGLUT3), and Slc1a2 (EAAT2) in isolated HEPs. j Representative immunofluorescence staining of VGLUT3 and EAAT2 in liver after in situ closed perfusion. Scale bar, 50 μm. Statistical comparisons were made using one-way ANOVA with Tukey’s multiple comparisons test. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.