Fig. 9
MiR-124-3p increases GLT1 expression by suppressing GLT1-inhibiting miRs in astrocytes. Representative GLT1 immunoblot (a) and quantification (b) following transfection of individual miRs (miR-124, miR-132, or miR-218) or co-transfection of miR-124-3p/miR-132 or miR-124-3p/miR-218 mimics in astrocyte cultures. n = 6–8 independent transfections per condition. P-values were determined from one-way ANOVA and post hoc Tukey’s test. Levels of miR-132 and miR-218 (c) and pri-miR-132 and pri-miR-218 (d) in astrocyte cultures following miR-124-3p transfection. n = 9 independent transfections per condition. β-actin mRNA was used as endogenous control for quantification of pri-miR-132. P-values were determined using one-way ANOVA and post hoc Tukey’s test. e Pie chart showing functional categories of genes that are expressed in astrocytes and are also predicted miR-124-binding targets. TargetScan was used to identify putative miR-124-3p target mRNAs. Ingenuity pathway analysis (Qiagen) was used for characterizing functional categories. Only genes that are expressed in both human and mouse astrocytes (FPKM > 10, mouse and human42 and have at least two predicted miR-124-3p-binding sites were included in generating the pie chart. f Representative Creb1 immunoblot from primary astrocytes following transfection with miR-124-3p and miR-124 antisense. The data was presented in the box and whisker plot with defined elements, median (center line), upper and lower quartiles (bounds of box), and highest and lowest values (whiskers)
