XB-IMG-130528
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Figure 2. Co(III)-Ebox binds target proteins to alleviate transcriptional repression in metastatic MDA-MB-231 cells.(A) E-cadherin reporter gene activity in MDA-MB-231 cells treated with Co(III)-Ebox from 0 to 40 nM. Data are represented as means ± s.e.m., n = 4. T-tests determined statistical significance from controls of 40 nM (P = 0.0489) (*). (B) E-cadherin reporter gene activity in MDA-MB-231 cells treated with 35 nM Co(III)-sb, Ebox, Co(III)-EboxMut or Co(III)-Ebox. Data are represented as means ± s.e.m., n = 4. T-tests determined statistical significance from controls of Co(III)-EboxMut (P = 0.0290) and Co(III)-Ebox (P = 0.0001) (*). Co(III)-Ebox to be significantly different from Co(III)-EboxMut (P = 0.0008) (**). (C) Luciferase reporter gene activity in MDA-MB-231 cells with EcadMut-luc reporter treated with 35 nM Co(III)-sb, Ebox, Co(III)-EboxMut or Co(III)-Ebox. Data are represented as means ± s.e.m., n = 4. T-tests determined no statistical significance between the means. (D) MDA-MB-231 cell viability after treatment with Co(III)-Ebox at indicated concentrations after 24 h. Data are represented as means ± s.e.m., n = 3. T-tests determined no statistical significance between the means. (E) Inhibition of Snail DNA binding in MDA-MB-231 cells after 24 h with 0 to 40 nM Co(III)-Ebox. A representative EMSA of triplicate samples is shown with Western blots of Snai1 and Actin. (F) A representative EMSA of MDA-MB-231 cells treated with shRNA targeting Snai1 with Western blots of Snai1 and Actin. (G) Quantification of the percentage of Slug probe bound to Snai1 in (E) by using a STORM 680 phosphoimager. Data are represented as the means ± s.e.m., n = 3. Image published in: Harney AS et al. (2012) Harney et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |