Fig. 8
Identification of miRs that inhibit GLT1 protein expression in astrocytes. a Schematic diagram of the human and mouse slc1a2 3′ UTR with predicted binding sites for miR-218 and miR-132. TargetScanMouse was used for the slc1a2 sequence analysis. Each arrow points to one binding site. b Expression levels of miR-218 and miR-132 in astrocytes in vitro (cultured astrocytes alone) and in vivo (at P7, P14, P21, and P70). Representative GLT1 immunoblots (c), quantification of GLT1 protein levels (d), and quantification by qPCR of glt1 mRNA levels (e) from cultured primary astrocytes following transfection of miR-132 or miR-218 mimics, miR-132 or miR-218 antisense (A/S), and miR-132 or miR-218 along with their respective antisense. n = 6–8 independent experiments per condition. P-values were determined using one-way ANOVA and post hoc Tukey’s test. n.s.: not significant; f Wild-type (WT) and miR-132 mutant (MT) GLT1 3′ UTR luciferase activity in HEK 293 cells following miR-132 transfection. g Wild type (WT) and miR-218 mutant (MT) GLT1 3′ UTR luciferase activity in HEK 293 cells following miR-218 transfection. n = 3 independent experiments with six replicates per experiment per condition; one-way ANOVA and post hoc Tukey’s test; 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)
