Fig. 5. JWA regulates GLT-1 expression via ERK/MAPK and PI3K/Akt signaling pathways.

a C8D1A cells were transfected with RFP-con or RFP-JWA. After 48 h, immunofluorescence for GLT-1 (green) was immunostained with anti-GLT-1 antibody. Scale bars = 200 μm. b C8D1A cells were transfected with si-con or si-JWA for various concentration (50 pmol or 100 pmol), followed by treatment with or without MPP⁺ 100 µM for 24 h. Expression levels of JWA and 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). c [³H]-Glutamate uptake assay was performed after C8D1A cells were transfected with si-JWA or Flag-JWA (n = 3). d C8D1A cells were transfected with si-JWA or Flag-JWA, followed by western blotting analysis to detect JWA, GLAST, GLT-1, phospho- and total-ERK, P38, JNK, and Akt proteins. The quantification for analysis of JWA, GLT-1, p-ERK, p-Akt protein (n = 3). Data are presented as mean ± SEM, one-way ANOVA, *P < 0.05, **P < 0.01