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

. 2019 Oct 28;10:4907. doi: 10.1038/s41467-019-12812-3

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

© The Author(s) 2019

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

Fig. 4 — Transcriptional enhancer function of TF regions. a Average plot of 30-way vertebrate Phastcons scores at distal H3K27ac and TF regions. b Average plot of Phastcons scores at distal regions co-occupied by the indicated number of TFs, or by distal heart H3K27ac regions that do not overlap TF regions. c Average Phastcons scores in distal regions (center ± 150 bp) co-occupied by the indicated number of TFs with or without heart H3K27ac in fetal or adult heart. Steel-Dwass: ****P < 0.0001; ***P < 0.001; **P < 0.01; NS, not significant, mean values represented by dot (black). d–f Individual enhancer activity in murine cardiomyocytes was measured using an AAV9-based assay. Vector construct (d) contained minimal hsp68 promoter driving mCherry, with enhancer element (Enh) located in the 3′ UTR. U6:2xBroccoli was used to normalize transduction efficiency. Twelve enhancers belonging to three groups (H3K27ac⁺ TF^–; H3K27ac⁺ ≥5TF⁺; H3K27ac^– ≥5TF⁺) were individually tested in vivo. Normalized enhancer activity was compared to empty vector (AAV9;Hsp68:mCherry-no enhancer; n = 9) using Dunnett’s test. Error bars represent s.d. e Representative dark field and fluorescent wholemount images of adult (P28) mouse hearts showing enhancer activity for each group of cardiac enhancers. g, h AAV-based massively parallel reporter assay measurement of the activity of 1982 candidate enhancers and negative controls in cardiomyocytes in vivo. AAV library was delivered at P0 and measured in ventricles at P7. g Activity, expressed as enhancer reads in RNA normalized to enhancer abundance in AAV genomic DNA, was plotted versus region activity rank. Relative rank of individual regions of each class is represented by vertical lines below plot. 5% false discovery rate (FDR) threshold (dotted line) indicates the 95th percentile of ESC H3K27ac negative control region activity. h Relative activity of each class of regions tested is summarized displaying mean (line) and 95th confidence interval (box) with s.d. (whiskers). Heatmap above violin plot shows statistical significance of inter-group comparisons (Steel-Dwass). Source data for panels c, e, g are provided as a Source Data file