Fig. 1: Inducible deletion of astrocytic GLUT1 in adult mice.
From: Astrocytic GLUT1 deletion in adult mice enhances glucose metabolism and resilience to stroke
a Generation of GLUT1fl/fl;GLASTCreERT2/+ (GLUT1 cKO) mice and littermate controls (GLUT1fl/fl;GLAST+/+). b Tamoxifen injections over 5 consecutive days in 8- to 10-week-old mice, with analyses conducted 60 days post-injection. c, d Western blot of GLUT1 (45 kDa) in capillary-depleted brain tissue shows a 48 ± 5% reduction in cKO mice (n = 4) compared to controls (n = 5, p = 0.0005, two-sided unpaired t-test). Vinculin served as loading control. Immunolabeling for GLUT1 and glutamine synthetase (GS) in cortex (e) and hippocampus (f) shows a marked reduction of GLUT1 signal in cKO brain sections (bottom panels). White arrows highlight GLUT1-depleted astrocytes, while capillaries (arrowheads) retain GLUT1 expression. Observed in four mice per genotype. g Cre-dependent AAV (PHP.eB-hGFAP-DIO-EGFPL10a) injected intravenously into GLUT1fl/fl;GLASTCreERT2/+ mice (cKO) and GLUT1+/+;GLASTCreERT2/+ mice (as controls) enabled astrocyte-specific EGFP-L10 expression for translating ribosome affinity purification (TRAP). Tamoxifen was administered 7 days post-AAV injection, and TRAP RNA was collected 60 days later. h Immunostaining confirms widespread EGFP-L10 expression (anti-GFP) in astrocytes (anti-S100β) using the approach in (g), observed in two mice per genotype. i qPCR analysis of astrocytic Gfap polysome-associated RNA following TRAP shows an 8.5-fold enrichment in immunoprecipitated (IP) samples vs. input in control (n = 5) and cKO mice (n = 5, p = 0.9873, two-sided unpaired t-test). j qPCR analysis of GLUT1 (Slc2a1) polysome-associated RNA in IP samples shows a 69 ± 5% reduction in cKO mice (n = 5) vs. controls (n = 5, p < 0.0001, two-sided unpaired t-test). k RNA-seq reads (counts per million, CPM) from exons 3–8 of Slc2a1 show a 68 ± 4% reduction in cKO cortical IP samples (n = 4) vs. controls (n = 3, p < 0.0001, two-sided unpaired t test). l RNA-seq analysis of glucose and monocarboxylate transporters reveals no significant differences between genotypes (n = 3 vs. 4, p > 0.85 for all genes, two-way Anova with Šídák’s multiple comparisons test). Data are presented as scatter dot plots with mean ± SEM. Source data are provided as a Source Data file.
