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. 2017 Oct 23;7:13856. doi: 10.1038/s41598-017-14364-2

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Suppression of CD44 decreases the expression of Ki67, and the capacities for mammosphere formation and tumorgenesis. (A) Analysis of CD44 mRNA levels following siRNA treatment. The mRNA expression decreased to 0.41 after CD44 siRNA treatment compared with control, Data represent means ± SD (n = 3), ****P < 0.0001. (B) Immunofluorescence images showing the expression of CD44, Ki67 in cells treated with control siRNA or CD44 siRNA, scale bars: 20 μm. (C) MDA-MB-231 cells transfected by control siRNA, CD44 siRNA or ALDH1 siRNA growing as non-adherent mammospheres after 10 d of mammosphere cultivation. Scale bar, 100 μm. (D) MDA-MB-231cells are tumorigenic while cells transfected by CD44 siRNA or ALDH1 siRNA could not form tumor with amount of 4 × 10⁶ in BALB/c nude mice after two weeks. All the experiments were performed in triplicate.