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. 2018 Sep 4;20:105. doi: 10.1186/s13058-018-1020-0

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© The Author(s). 2018

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. 5 — Analysis of breast cancer stemlike phenotype in vMCF-7^∆Raf1 1GX cells with abrogated NOTCH3 expression. a Immunofluorescence analysis showing representative images of vMCF-7^∆Raf1 1GX and vMCF-7^∆Raf1 1GX/CRISPR-NOTCH3 cells stained in green with a CD44 polyclonal antibody and in red with a CD24 monoclonal antibody. Nuclei were stained in blue with 4′,6-diamidino-2-phenylindole (DAPI). b Graph showing the average of cells expressing a CD44⁺/CD24⁻ phenotype from three independent experiments (± SD). c Immunofluorescence analysis showing representative images of vMCF-7^∆Raf1 1GX and vMCF-7^∆Raf1 1GX/CRISPR-NOTCH3 cells stained in red with an estrogen receptor alpha (ERα) monoclonal antibody. Nuclei were stained in blue with DAPI. d Graph showing the average of ERα-positive cells from three independent experiments (± SD). e Fluorescence-activated cell sorting analysis showing aldehyde dehydrogenase 1 (ALDH1) activity in vMCF-7^∆Raf1 1GX and vMCF-7^∆Raf1 1GX/CRISPR-NOTCH3 cells. Samples treated with the ALDEOFLUOR inhibitor N,N-diethylaminobenzaldehyde (DEAB) were used as a negative control. f Graph showing the average of ALDH1⁺ cells from three independent experiments (± SD)