Fig. 6. Transcription factor CREB is responsible for JWA-mediated GLT-1 expression.

a, b C8D1A cells were transfected with si-JWA or Flag-JWA. After 48 h, the mRNA levels of GLAST and 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 = 3). c C8D1A cells were transfected with si-JWA or Flag-JWA, followed by western blotting analysis to detect transcription factor p-P65, YY1, phospho- and total-CREB proteins (n = 3). d, e C8D1A cells were incubated with 50 µM of U0126 and/or 20 µM of LY for 3 h after transfect with Flag-con or Flag-JWA for 48 h, followed by western blotting analysis to detect p-ERK, p-Akt, p-CREB and GLT-1 (n = 3). [³H]-Glutamate uptake assay was also performed (n = 3). f C8D1A cells were co-transfected with either Flag-con or Flag-JWA, together with the si-con or si-CREB. After 48 h, expression levels of GLT-1 were detected by western blotting, GAPDH was used as a loading control to confirm that equal amounts of protein were loaded in each line (n = 3). g SH-SY5Y cells were treated for 24 h with astrocyte CM from astrocytes transfected with si-con or from astrocytes transfected with si-JWA. After the treatment, SH-SY5Y cells were exposed to MPP⁺ (50 μM or 100 μM) for 24 h. The expression levels of TH were detected by western blotting, 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