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 13;8:9072. doi: 10.1038/s41598-018-27280-w

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

© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

PER2 represses p65/p50-mediated Slc9a3r1 transcription. (a) Schematic representation of luciferase reporter constructs that contain tandemly repeated E-boxes (4 × E-boxes::Luc), ROREs (4 × ROREs::Luc), D-sites (4 × D-sites::Luc), or PPREs (4 × PPREs::Luc). (b) Effects of clock gene products on the transcriptional activities of 4 × E-boxes::Luc, 4 × ROREs::Luc, 4 × D-sites::Luc, 4 × PPREs::Luc, and Slc9a3r1(−200/+168)::Luc. Control groups were transfected with empty vector (pcDNA3.1) instead of expression plasmids. Presence (+) or absence (−) of expression plasmids (0.5 µg each) is denoted. Each value represents the mean ± s.e.m. (n = 6). **P < 0.01, significantly different between the two groups. (ANOVA with Tukey-Kramer post hoc test). (c) Cells were transfected with Slc9a3r1(−200/+168)::Luc and Slc9a3r1 (+35/+168)::Luc in the absence or presence of Per2 expression plasmids. Each value represents the mean with s.e.m. (n = 6). **P < 0.01, significant difference between the two groups (F_11,60 = 58.935; P < 0.001; ANOVA with Tukey-Kramer post hoc test). (d) Enhancement of the transcriptional activity of Slc9a3r1(−200/+168)::Luc in Per2 mutant (Per2^m/m) cells. Mouse embryonic fibroblasts (MEFs) were prepared from wild-type and Per2^m/m mice. Cells were transfected with Slc9a3r1(−200/+168)::Luc or Slc9a3r1 (+35/+168)::Luc. Each value represents the mean with s.e.m. (n = 6). **P < 0.01, significant difference between the two groups (F_3,20 = 27.200; P < 0.001; ANOVA with Tukey-Kramer post hoc test). (e) The down-regulation of PER2 by miRNA enhances the transcriptional activity of Slc9a3r1(−200/+168)::Luc. The left panel shows a representative photograph of the immunoblot analysis of the PER2 protein in miRNA-expressing vector-transfected cells. Each value represents the mean with s.e.m. (n = 6). **P < 0.01, significant difference between the two groups (F_3,20 = 516.818; P < 0.001; ANOVA with Tukey-Kramer post hoc test). (f) Cells were transfected with luciferase-reporter vectors containing the consensus NF-кB response element (NF-κB-RE::Luc), Slc9a3r1 (−200/+168)::Luc, or Slc9a3r1 (+35/+168)::Luc in the absence or presence of expression plasmids encoding p65, p50, or PER2. Each value represents the mean with s.e.m. (n = 6). **P < 0.01, significantly different from the control group; ^##P < 0.01, significantly different from the p65/p50-transfected group (F_3,20 = 127.370; P < 0.001 for Slc9a3r1(−200/+168)::Luc; F_3,20 = 68.062; P < 0.001 for NF-κB-RE::Luc; ANOVA with Tukey-Kramer post hoc test). Full-size images of western blotting are presented in Supplementary Figs 10 and 11.