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. Author manuscript; available in PMC: 2017 Dec 1.
Published in final edited form as: Exp Neurol. 2016 Oct 8;286:137–146. doi: 10.1016/j.expneurol.2016.10.003

Figure 1. Levels of miR-21 transcripts at different time points in rats after SE.

Figure 1

(A) A schematic diagram of miR-21 biogenesis. MiR-21 is transcribed as a long primary transcript (~3400 bp), pri-miR-21, from its own promoter. Drosha cleaves pri-miR-21 into a precursor transcript (~ 70 bp), pre-miR-21. Dicer further cleaves pre-mi-21 into short strands of ~22 nt, containing guide/mature miR-21 (green) and passenger miR-21 (miR-21*) (magenta). (B–C) qRT-PCR showing changes of pri-, pre-, and mature miR-21 levels at 4, 12, and 48 h after SE. Both pri- and mature miR-21 levels show significant increase only at 12 h and 48 h, respectively (B & D), whereas pre-miR-21 levels are elevated at all time points (C). Relative fold change is normalized to PPIA (cyclophilin) mRNA expression for pri-miR-21 and to 4.5S (ribonuclear small RNA) expression for pre-miR-21 and for mature miR-21. Shown are mean±SEM. Kruskal-Wallis test, *p<0.05, **p<0.01, *** p<0.001. (E & F) Small RNA northern blots detecting pre-miR-21 and mature miR-21 at 12 and 48 h post SE. The blots used LNA probe that detected both pre-miR-21 (red arrowhead) and mature miR-21 (circle). At 12 h time point, the intensity of the pre-miR-21 bands in the SE groups was stronger than that in the control group, but the intensities of the mature miR-21 bands were comparable between the SE and control groups (E). At 48 h time point, the levels of pre-miR-21 and mature miR-21 in the SE group were higher than that in the control group (F). (G & H) Quantification for northern blots. L= ladder for RNA sizes in nucleotides. U6 serves as a loading control. For pri-miR-21: 4h (Control N=10, SE N=12), 12h (Control N=7, SE N=8), 48h (Control N =11, SE N=10). For pre-miR-21: 4h (Control N= 7, SE N=11), 12h (Control N=7, SE N=9), 48h (Control N=5, SE N=6). For mature miR-21: 4h (Control N= 6, SE N=6), 12h (Control N=4, SE N=5), 48h (Control N=6, SE N=6).