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. 2014 Dec 9;16:482. doi: 10.1186/s13058-014-0482-y

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© Hill et al.; licensee BioMed Central. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Cellular response to ETP-45658. (A) Proliferation assay for each indicated cell line treated with serial dilutions of ETP-45658 or reference compound PI-103. Seventy-two hours post treatment, MTT assays were conducted. (B) Dose- and time-dependent analysis of Ser473 phosphorylation of AKT phosphorylation by 100nM ETP-45658 or PI-103 treatment. MCF-7 or U2OS cells were treated with the indicated concentrations (μM) of ETP-45658 (or PI-103) for four hours, then total protein was extracted and immunoblotted. For nuclear FOXO analysis, nuclear protein fractions were collected four hours post ETP-45658 (or PI-103) treatment. Nuclear FOXO3a levels were measure by immunoblotting. Nuclear fractions were confirmed by probing for lamin A/C. (C) FACS profile of MCF-7 or U2OS following 100 nM ETP-45658 treatment (20,000 events scored, N = 5). (D) Heat map for the top 100 differentially expressed genes based on hierarchical clustering using Euclidean distance metric. Red and green indicate genes that induced or repressed respectively by ETP-45658 or PI-103. Black indicates no change (N = 4). (E) Venn diagram of upregulated genes after ETP-45658 or PI-103 treatment (100nM). (F) Venn diagram of downregulated genes after ETP-45658 or PI-103 exposure. Numbers indicate number of genes that are exclusive to that particular category.