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. 2018 Nov 23;7(11):91. doi: 10.1038/s41389-018-0101-3

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© 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/.

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Fig. 5 — a, b Comparative analysis of Ets1 expression levels in transcripts and protein between WT and CRE-deleted MDA-MB-231 (ΔCRE) cells determined by qRT-PCR and Immuno-blot. Transcript levels were normalized against Hprt and actin served as loading controls, respectively. Data are presented as mean ± SD. Two-way ANOVA with Bonferroni post-tests showed a significant difference of Ets1 expression. c Heatmap of RNA-sequencing data showing differentially expressed genes (DEGs) in WT and ΔCRE MDA-MB-231 cells. d GO-term enrichment using DAVID “Biological functions” category. Shown are the top five up and down-regulated functions based on –log2(p value). e Comparison of expression levels of Ets1 target genes between WT and ΔCRE cells. f ChIP-PCR was performed to detect Ets1 enrichment on the promoter regions of Eng and Mmp14 genes in MDA-MB-231 cells 2 h after stimulation. Fold enrichment compared with input is shown. Data are representative of two individual experiments. Bars represent averages from triplicate PCR reactions as mean ± SD. One-way ANOVA with Bonferroni correction indicated a significant enrichment of Ets1; *p < 0.05; **p < 0.01; ***p < 0.001. g Scatterplots and nonparametric Spearman’s rank correlation analysis with corresponding P-values correlation analysis from TCGA. Correlation between Ets1 and target genes (Eng and Mmp14) in mRNA levels. Each symbol represents an individual BRCA sample. h Scatterplots and nonparametric Spearman’s rank correlation analysis with corresponding p-values correlation analysis from CCLE. Correlation between Ets1 and target genes (Eng and Mmp14) in mRNA levels. Each symbol represents an individual human breast cancer cell line. i Cells were stained with crystal violet and representative images were obtained from in vitro invasion assay using 10% FBS as chemoattractant. Cells were stained with crystal violet. Scale bar: 100 m. j Representative images (left) and numbers of metastasis nodules (right) of mouse lungs obtained from 6 weeks after tail vein injections of WT or ΔCRE cells (each 5 × 10⁵) into eight female nude mice, respectively. i, j Data are presented as mean ± SD. One-way ANOVA with Bonferroni correction indicated a significant difference between WT and ΔCRE cells. a, b, e, f, i, j Data shown are representative of more than two independent experiments with similar results. *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t-test)