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

. 2016 Nov 4;7:13287. doi: 10.1038/ncomms13287

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

Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

PMC Copyright notice

(a) A scheme of vectors expressing different GAS5 truncated transcript. The inserted length of each transcript is indicated and the blue square indicates the position of validated GR-binding sequence. 1-651mut represents the GR-binding sequence-mutated construct. The right panel shows the mRNA level of NANOG and NODAL in different treatment groups. **P<0.01, t-test, n=3. (b) The left panel shows the scheme of constructing NODAL promoter reporter. The right panel shows the luciferase activities of NODAL promoter reporter in hESCs with indicated treatment. NC represents NODAL promoter reporter transfected hESCs treated with empty vector, t-test, n=3. (c) The nuclear-plasma separation assay of GAS5 overexpressed or knockdown hESCs are shown in the middle panel using qPCR. Respective FISH images are shown in the left panel, and the GAS5 expression of analysed using total RNAs are shown in the right panel (n=3). NUCL represents nuclear hESC extracts, CYTO represents cytoplasm hESC extracts. Scale bar, 20 μm. All groups were compared with NC or NC-NUCL group respectively. **P<0.01, t-test, n=3. (d) A scheme of MS2-mediated pulldown of GAS5 and its binding protein. (e) The silver staining of pull-down products (n=2), the red triangle indicates the antibody band, whereas black triangle indicates the bands and position that is cutoff for mass spectrometry analysis shown in the right panel. (f) Western blot shows the AGO2 expression in different pulldown groups as indicated by the black triangle (n=2). The red triangle indicates the antibody heavy chain bands, which were used in pull-down experiment and react with the secondary antibody. (g) Confocal microscopy of AGO2 and GAS5 transcript. Scale bar, 100 μm. (h) RNA immune-precipitation analysis of the RNA levels of precipitated GAS5, linc-ROR, H19 using indicated protein antibodies by qPCR. **P<0.01, t-test, n=3. (i) AGO2 competing assay using AGO2 antibody-mediated RNA-IP. RNA levels of precipitated NODAL, linc-ROR, H19 are tested using qPCR. **P<0.01, t-test, n=2. Error bars represent s.d. of the indicated experiment replicates. RNA level of β-actin served as internal reference for qPCR. See also Supplementary Fig. 5.