Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Jun 11;11:171. doi: 10.3389/fnmol.2018.00171

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2018 Zampa, Bicker and Schratt.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Design and validation of functionality and subcellular localization of pre-miR-134 chimeras. (A) Graphic representation of pre-miR-134 chimeras tested for subcellular localization. The pre-miR-134 terminal loop was substituted with either the one from pre-miR-150 (pre-miR-134L150) or the one from pre-miR-181a (pre-miR134L181a). (B) Representative pictures of developing hippocampal neurons transfected with in vitro synthesized Cy3-labeled pre-miRNAs described in (A). Upper panel: merged images of Cy3-pre-miRNAs (green) and MAP2 antibody staining (red). Lower panel: high contrast black-and-white image of the Cy3-pre-miRNA signal only. Arrows point to dendritic pre-miR-134 granules (scale bar: 20 μm). (C,D) Quantification of dendritic pre-miRNA granules from multiple neurons shown in (B). Values represent the average percentage of dendritically localized (C) or the total number/cell (D) of Cy3-pre-miRNA granules ± SD. n = 40 cells analyzed per data point from n = 3 independent biological replicates. One-way ANOVA and Tukey’s, ***P < 0.001. (E) Luciferase reporter gene assays performed using plasmids containing either 3xmiR-134 perfect binding sites (3x-miR-134-PBS) or the corresponding reverse sequence (3x-miR-134-REV) in the firefly luciferase 3′UTR. Firefly reporters were co-transfected with the indicated pre-miRNA expression vectors (or empty pcDNA3) and Renilla luciferase expression was used for internal normalization. Firefly/Renilla ratios were calculated for miR-134 reporters and subsequently normalized to the ratios obtained with the pmirGLO empty vector. The condition co-transfected with empty pcDNA3 was set to one (not shown). Values represent the mean normalized luciferase activity ± SD of n = 4 independent biological replicates. One-sample t-test, *P < 0.05.