Fig. 2: Glutamate accumulation in perivenous hepatocytes through vesicular glutamate transporter 3 after 2-week EtOH intake and its release by binge drinking.
a Glutamate concentrations in serum and liver from Pair-fed and EtOH-fed WT mice with/without binge drinking (n = 8/group). b RNA-sequencing analysis of the whole liver tissues from WT mice fed with isocaloric (Pair) or EtOH diet for 2 weeks (n = 3/group). Graphical representation and heatmaps about pathways of glutamate production and transportation. c Relative mRNA expression of glutamate transporters according to the liver zonation in single-cell RNA (scRNA)-sequencing of WT mouse HEPs (GSE84498). d Representative immunofluorescence staining of VGLUT3, EAAT2, OAT, and ALDH4A1 in liver sections merged with DAPI. Central vein (CV) and portal triad (PT). Scale bar, 50 μm. e Western blot analysis in isolated HEPs from WT mice. f Relative mRNA expression of Aldh4a1, Oat, Slc1a2, and Slc17a8 in the whole liver tissues from EtOH-fed mice and EtOH-treated HEPs compared to control (n = 4/group). g Immunostaining of VGLUT3 after liver tissue expansion by the epitope-preserving magnified analysis of proteome protocol. Scale bar, 100 μm. h Schematic protocol of isolating microvesicles (MVs) by stepwise centrifugation. i, j Representative Western blot analysis and glutamate concentrations in isolated MVs from Pair- or EtOH-fed WT mice (n = 6/group). Statistical comparisons were made using one-way ANOVA with Tukey’s multiple comparisons test (a) or two-tailed unpaired t-test (f, j). p < 0.05 was considered statistically significant. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.
