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. 2015 Dec 15;14:209. doi: 10.1186/s12943-015-0478-y

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© French et al. 2015

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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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Fig. 4 — Nuclear cFLIP promotes Wnt-signalling (a) Western blotting for cFLIP and beta-catenin performed on cytoplasmic protein extracts of MCF-7 and MDA-MB-231 cell lines transfected with siRNA targeting cFLIP or a scrambled control (loading control alpha-tubulin) (b) Immunofluorescence for cFLIP and beta catenin of MDA-MB-231 cells transfected with siRNA targeting cFLIP or a scrambled control (red = cFLIP, green = beta catenin, blue = DAPI) (c) qPCR analysis for mRNA levels of cFLIP and Axin2 in MCF-7 and MDA-MB-231 cells transfected with siRNA targeting cFLIP or a scrambled control and cultured in the presence of 10 ng/ml Wnt3a for 48 h. Gene expression is expressed as relative to the untreated control. (d) TOPFlash luciferase reporter assay of MCF-7 cells transfected with overexpression constructs; mock (empty), cytoplasmic-localised cFLIP and nuclear-localised cFLIP and cultured in the presence of 10 ng/ml Wnt3a for 48 h. Luciferase output is expressed as relative to that of the mock-transfected cells. (*p < 0.05, paired t-test, error bars = SEM; all graphs represent means of at least 3 independent experiments)