Fig. 4. Effect of JWA on astrocyte reactivity and GLT-1 expression in primary cultured astrocytes.

a After primary cultured astrocytes were isolated from JWA WT and JWA CKO, immunofluorescence for GFAP (red) was immunostained with anti-GFAP antibody. Scale bars = 20 μm. b The primary cultured astrocytes were transfected with si-JWA or Flag-JWA. After 48 h, the mRNA levels of GLT-1 were measured by quantitative real-time PCR. GAPDH was used as a control to normalize the differences in the amount of total RNA in each sample (n = 6). c [³H]-Glutamate uptake assay was performed after primary astrocytes were transfected with si-JWA or Flag-JWA (n = 3). d Representative western blotting results were detected using anti-GLT-1 antibody. GAPDH was used as a loading control to confirm that equal amounts of protein were loaded in each line (n = 3). Data are presented as mean ± SEM, one-way ANOVA, *P < 0.05, **P < 0.01