Fig 2: The percentage of GLT-1-positive maturing (O4-positive) OLGs is transiently reduced in male Glt-1 Plp1icKO brains at 4 weeks of age. (A,C) Graphs depicting the percentage of maturing (O4-positive) OLGs with surface expression of GLT-1, as determined by flow cytometry, at 4 (A) and 6 (C) weeks of age. Glt-1 Plp1icKO and Wt (Plp1CreERT-negative littermate) mice were treated as depicted in Figure 1A and analyzed at 4 and 6 weeks of age. Dots represent individual samples (animals), horizontal lines indicate means, error bars are depicted as SEM. *p = 0.05, multiple comparisons, two-way ANOVA with post-hoc Tukey’s test; there is no significant interaction; individual p-values are depicted for p > 0.05. The inset in panel (A) depicts a representative histogram displaying relative fluorescence intensity (x-axis) vs. event number (y-axis) gated for O4 positivity; overlaid histogram subtraction (GLT-1 histogram—IgG isotype control histogram) was used to determine the percentage of GLT-1-positive maturing (O4-positive) OLGs. (B) Representative genomic PCR results for brain tissue taken from Glt-1 Plp1icKO and Wt (Plp1CreERT-negative littermate) female and male mice.

Fig 3: Conditional deletion of Glt-1 from maturing OLGs does not significantly affect myelin gene expression in the corpus callosum of female and male mice. (A,B) Graphs depicting mRNA levels for the myelin genes genes myelin basic protein (Mbp), myelin oligodendrocyte glycoprotein (Mog), and proteolipid protein (Plp1) in female (A) and male (B) corpus callosum as determined by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). (C) Graph depicting PLP protein levels as determined by Western blot analysis. Glt-1 Plp1icKO and Wt (Plp1CreERT-negative littermate) mice were treated as depicted in Figure 1A and analyzed at 4 weeks of age. Dots represent individual samples (animals), horizontal lines indicate means, error bars are depicted as SEM; individual p-values are depicted for p > 0.05, unpaired t-test (two-tailed).

Fig 4: Human NPCs differentiate into mature neurons. Immunofluorescence staining of differentiated neurons derived from human dorsal NPCs (1323–2 line, day 35 after differentiation) for mature cortical neuronal markers expressed in the nucleus (BRN2, TBR1, NeuN) and cytoplasm (MAP2), glial markers (SOX9, GFAP, GLT1), and dorsal forebrain marker (FOXG1). Nuclei stained with DAPI, shown as an overlay over brightfield images. The merge is an overlay of the neuronal and glial markers. Scale bar 100 µm. DAPI 4,6'-diamino-2-phenylindole

Fig 5: Hippocampal GLT-1 and NR1 expression 3 days after SE with MyD88 inhibition. Sections from the hippocampi of mice in the CP group (A1–A3) and MIP group (B1–B3) 3 days after SE with GLT-1 immunoreactivity in astrocytes and neuronal processes. (A4, B4) Higher magnification of the boxes in (A3) and (B3). (C) Comparison of the numbers of GFAP/GLT-1 double-labeled cells in the DG, CA1, and CA3 between the CP and MIP groups (means ± SEM, n = 3). *p < 0.05 versus the CP group; **p < 0.01 versus the CP group. Independent samples t tests were performed. (D1) Immunoblots of NR1, NR2a, and NR2b for the control, CP, and MIP groups. (D2–D4) Comparison of NR1, NR2a, and NR2b levels among the above groups (calibrated to ß-actin). *p < 0.05; ***p < 0.001 between groups. One-way ANOVA followed by Tukey’s test. Scale bars: (A1–A3, B1–B3) 100 µm; (A4, B4) 50 µm