Journal: PLoS ONE

Article Title: Transcription Factor Ets1 Cooperates with Estrogen Receptor α to Stimulate Estradiol-Dependent Growth in Breast Cancer Cells and Tumors

doi: 10.1371/journal.pone.0068815

Figure Lengend Snippet: A. Breast cell lines were analyzed by Western blot for ERα and Ets1 with tubulin as a loading control. B. ER-positive BT-474, MCF7, T47D and ZR-75-1 cells were analyzed by qRT-PCR for Ets1 transcript levels. Data represent mean ± standard deviation. Ets1 transcript levels are statistically significantly lower in BT-474, MCF7, T-47D and ZR-75-1 cells compared to MCF10A cells by ANOVA with Tukey’s post-test analysis p<0.001.

Article Snippet: MCF7 cells were purchased from the American Type Culture Collection.

Techniques: Western Blot, Control, Quantitative RT-PCR, Standard Deviation

Journal: PLoS ONE

Article Title: Transcription Factor Ets1 Cooperates with Estrogen Receptor α to Stimulate Estradiol-Dependent Growth in Breast Cancer Cells and Tumors

doi: 10.1371/journal.pone.0068815

Figure Lengend Snippet: Colonies formed as a function of Ets1 expression.

Article Snippet: MCF7 cells were purchased from the American Type Culture Collection.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Transcription Factor Ets1 Cooperates with Estrogen Receptor α to Stimulate Estradiol-Dependent Growth in Breast Cancer Cells and Tumors

doi: 10.1371/journal.pone.0068815

Figure Lengend Snippet: A. Whole cell lysates from MCF7 control and Ets1 stable transformants with equal amounts of total protein were immunoprecipitated with an anti-HA.11 antibody and analyzed by Western blot for Ets1expression. Corresponding whole cell lysates were assessed by Western blot analysis for ERα with anti-tubulin as a loading control. B. Stable transformants were plated at a density of 2000 cells/well in phenol red free media with 5% charcoal dextran stripped serum. On Day 3, the wells were treated with either 0.1% ethanol vehicle, or 10 nM 17β-estradiol (E2). Cell growth was assessed over eight days using a resazurin-based assay. Data represent the mean ± SEM of two independent experiments. * Different from control or non-estradiol stimulated culture by ANOVA with Tukey’s post-test analysis p<0.05. # Different from control+E2 p<0.05. C. Control and Ets1-expressing stable transformants were plated at a density of 800 cells/well. Estradiol (10 nM) was added as indicated. After 13–14 days colonies containing ≥50 cells were counted. Data represent the mean ± SEM of five independent experiments. Significantly different from control (CTV, *) or control+estradiol (#) by ANOVA with Dunnett’s post test analysis p<0.05. D. Log phase control (CTV) and Ets1 stable transformants were resuspended (50,000 cells/well) in serum and phenol red free medium in 24-well BD migration and matrigel invasion chambers with 5% FBS as the chemoattractant. After 24 hours, the filters were scrubbed, fixed, stained and counted. Data represent the mean ± SEM of three independent experiments performed in triplicate. * Significantly different from control by ANOVA with Dunnett’s post test analysis, p<0.05.

Article Snippet: MCF7 cells were purchased from the American Type Culture Collection.

Techniques: Control, Immunoprecipitation, Western Blot, Resazurin Assay, Expressing, Migration, Staining

Journal: PLoS ONE

Article Title: Transcription Factor Ets1 Cooperates with Estrogen Receptor α to Stimulate Estradiol-Dependent Growth in Breast Cancer Cells and Tumors

doi: 10.1371/journal.pone.0068815

Figure Lengend Snippet: Female athymic nu/nu mice were ovariectomized and subcutaneously implanted with either an estradiol or control silastic pellet. After one week recovery, 2×10 6 cells of either control or Ets1-expressing stable cells (clonal isolate #1) were injected into the right mammary fat pad (n = 10/group). A. Tumor volume was calculated using the 3.14 x length(w 2 )/6. Values are mean ± SEM. Symbols indicate values that differ significantly from estradiol/MCF7 tumors by ANOVA with Bonferroni’s post-test analysis. * p<0.01, # p<0.001 day 34 to end. B. At end of study, tumors were excised and weighed. *** Significantly different from MCF7/estradiol p<0.001. C. Mice were imaged with the Xenogen Bioluminescent system after one week to assess tumor implantation and at the end of the study. Representative images for each group at constant signal intensity are shown. D. Representative images of hematoxylin and eosin stained tumor sections, Ki67(red), TUNEL (red) overlayed on DAPI (blue) staining in tumors. E. Ki67 stained sections from representative tumors (n = 7). Bars represent mean ± SEM of the average tumor staining. F. TUNEL stained sections from representative tumor sections (n = 7). Bars represent mean ± SEM of the average tumor staining. ** Significantly different by t-test p<0.05.

Article Snippet: MCF7 cells were purchased from the American Type Culture Collection.

Techniques: Control, Expressing, Injection, Tumor Implantation, Staining, TUNEL Assay

Journal: PLoS ONE

Article Title: Transcription Factor Ets1 Cooperates with Estrogen Receptor α to Stimulate Estradiol-Dependent Growth in Breast Cancer Cells and Tumors

doi: 10.1371/journal.pone.0068815

Figure Lengend Snippet: Ets1-expressing stable cell lines and MCF7 control cells were analyzed by qRT-PCR for expression of Cyclin D1 (A), PGR (B) and TFF1 (C). * Significantly different than control MCF7 cells by t-test, p<0.05.

Article Snippet: MCF7 cells were purchased from the American Type Culture Collection.

Techniques: Expressing, Stable Transfection, Control, Quantitative RT-PCR