Fig. 1

OSM down-regulates GLT-1 and GLAST expression in primary mouse cortical astrocytes. a Total mRNA purified from wild-type (C57BL/6J) mouse neonatal (P2) brain cortex as well as from cultured cortical astrocytes established from P2 brains was analyzed for expression of OSMR-β and gp130 mRNA by reverse transcriptase PCR; GAPDH primers were used as loading control. b Cortical astrocyte cultures were treated without or with OSM (10 ng/mL) for 2, 4, 8, 12, and 24 h and analyzed for GLT-1 and GLAST mRNA levels (gene expression normalized to HPRT1) by real-time PCR. Data are normalized to untreated controls and presented as mean ± SEM; n = 3, **p < 0.01, ***p < 0.001; one-way ANOVA using Bonferroni correction. c Cortical astrocytic cultures were treated without or with OSM (10 ng/mL) for 3, 5, and 24 h and were analyzed for GLAST proteins by Western blot. Relative densitometric analysis of GLAST proteins is shown in the lower panel. Data are presented as percentage of each respective ratio between optical density value of GLAST band intensity and optical density value of the matched α-tubulin (which served as the loading control) band intensity; n = 3, **p < 0.01, ***p < 0.001; one-way ANOVA