Figure 4.

PBMT upregulates GLT-1 expression by activating the PI3K/Akt/NF-κB signaling pathway in CORT-treated primary astrocytes. (a) CORT induced reduced viability of primary cultured astrocytes in dose-dependent manners as measured using the CCK-8 assay. The ordinate represents the percentage of cell survival compared with controls as measured by the CCK-8 assay. (b) Primary astrocytes were exposed to 200 μΜ corticosterone followed by irradiation with PBMT at 1 J/cm², 2 J/cm², or 4 J/cm², respectively. Cell viability was assessed by the CCK-8 assay after 24 h. (c) Western blot and quantification analysis of GLT-1 expression in 200 μΜ CORT-treated primary astrocytes. (d, e) Representative PCR and western blot and quantification analysis that PBMT increases GLT-1 mRNA and protein levels in a dose-dependent manner. (f) Representative immunofluorescent images of p65 (green) in primary astrocytes under the indicated treatments. Staining with DAPI (blue) to visualize the nucleus. Scale bar: 10 μm. (g, j) GLT-1 mRNA levels were detected by PCR stimulated with CORT and/or PBMT in the preference of API-2 (6 μΜ) and PDTC (8 μΜ) in primary astrocytes. (h, i) Representative western blot and quantification analysis of GLT-1 stimulated with CORT and/or PBMT in the preference of API-2 (6 μΜ) and PDTC (8 μΜ) in primary astrocytes. (k) Representative immunofluorescent images of GLT-1 (green) in astrocytes (red) under the indicated treatments. Staining with DAPI (blue) to visualize the nucleus. Scale bar: 20 μm. All the data represent mean ± SEM. ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. Significant differences were analyzed by the two-sided unpaired Student's t-test for two-group comparisons and one-way ANOVA followed by Tukey's post hoc test for multiple comparisons. CORT: primary astrocytes treated with corticosterone.