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 Jul 30;9:2980. doi: 10.1038/s41467-018-05367-2

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

Fig. 4 — EVL is a transcriptional target of ER. a Differential gene expression analysis of actin cytoskeletal regulators in ER+ vs. ER− breast tumors. Volcano plot of significance (−log₁₀ of p value) vs. log₂ fold change. Horizontal dashed line represents threshold for significance at p < 0.05. Vertical dashed lines represent threshold for positive or negative twofold change in gene expression. Insets are lists of genes that passed both thresholds in descending order of significance (lists were limited to five genes; full data analyses are presented in Supplementary Data 3). b Unsupervised clustering analysis of TCGA RNA-seq dataset. Heatmap, generated by row scaling, showing expression of genes in the ESR1 cluster. Tumor samples were classified into ER+ (black) and ER− (white) based on ER status (paired t test used to determine significance). c Analysis of ER binding by chromatin immunoprecipitation (ChIP). Genomic region surrounding EVLshowing ER-ChIP sequencing profile as SPMR (signal per million reads) trace of two independent ER-ChIP sequencing samples, generated by MACS2.0 and visualized by IGV (Integrative Genomics Viewer). Corresponding input profiles are shown at ×10 scale to clearly show the input DNA openness bias. d Fold enrichment of ER binding at four peaks found by ChIP-seq, in addition to positive (PGR) and negative controls. Fold enrichment is calculated as [DNA]-normalized ChIP/Input of 2^ΔΔCt. Values are means of data from four replicates; mean ± s.e.m.; p values were generated by paired t test comparing E2 treated to vehicle treated and E2 treated to negative control; ^†p = 0.009, ^††p = 0.007, ^†††p = 0.001, ^††††p = 0.001. e Analysis of the Regulation of EVL expression by ER. qPCR of EVL mRNA in MCF7 cells treated with corresponding drugs for 24 h, normalized to GAPDH. Fold change shown between treatment groups. Values are means of data from four independent experiments; mean ± s.e.m.; *p = 0.03, ***p = 0.0008 (unpaired t test). f Immunolabeling of MCF7 cells after corresponding drug treatment for 72 h. Bottom row shows EVL labeling. Scale bar is 10 µm. g Quantification of EVL levels in MCF7 cells after treatment with corresponding drugs for 72 h. Values are means of data pooled from two independent experiments; mean ± s.e.m. ^†p = 0.03, ^††p = 0.03, ^†††p = 0.04 (Welch’s t test)