Skip to main content
. Author manuscript; available in PMC: 2016 Mar 31.
Published in final edited form as: Oncogene. 2015 Jul 13;35(13):1643–1656. doi: 10.1038/onc.2015.226

Figure 2. Interaction of TOB1 with the signaling landscape of estrogen-dependent and estrogen-independent cells.

Figure 2

A. Comparison of steady state mRNA expression of 45 genes active in LCC1 and/or LCC9 in 9 estrogen-dependent and 8 estrogen-independent breast cancer cell lines (Table S4). Data obtained from Cancer Cell Line Encyclopedia [8] [4]. Unsupervised clustering was performed using MeV. The color scale represents mRNA levels with the darkest blue indicating the lowest level and the darkest yellow indicating the highest level. Red arrows indicate top 2 genes with most significant different expression between estrogen-dependent and –independent cell lines. B. Percentage of gene alteration in Breast Invasive Carcinoma from TCGA (1033 tumors). C-F, TOB1 is not sufficient to induce estrogen independence in MCF7 cells. TOB1-expression plasmid pLOC-TOB1, or vector control, and VSV-Gal vectors were packaged into HEK 293 cells to produce lentivirus containing TOB1. Cells were transduced with virus and selected by Blasticidin S (10 μg/ml for MCF7, 11 μg/mL for LCC1 and LCC9 cells). TOB1 protein level was detected by western blotting. Growth of TOB1-overexpressed cells was determined by CellTiter-Blue. C. Levels of TOB1 protein and p-TOB1 in TOB1-overexpressed MCF7 cells. Representative images of western blots are shown. D. Quantitation of protein levels shown in C. E. Growth of MCF7 cells expressing vector control DNA or TOB1 in the absence of estrogen. Cell growth measurements were determined with CellTiterBlue treatment. F. Proliferation of MCF7 cells expressing vector control or TOB1 in the presence of 1nM estradiol. TOB1 ORF1 and ORF2 were two constructs used to obtain enhanced TOB1 exogenous expression.