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. 2017 Jun 27;7:4263. doi: 10.1038/s41598-017-02800-2

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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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Rescue of BMI1 level as well as inhibiting endogenous miR-15a, miR-16 reduces miR-15a, miR-16 mediated DNA damage. Expression of BMI1, ϒ-H2AX, p-ATM, Ub-H2A and RNF8 were checked in MCF-7, MDAMB-231 cells transfected with anti-miR-15a, anti-miR-16 or both anti-miR-15a/16. Actin served as gel loading control. Blots were cropped to enhance the representation (A). Expression of BMI1, ϒ-H2AX, Ub-H2A, RNF8, p-CHK2, p-ATM were checked in MDAMB-231 cells co-transfected with miR-15a, miR-16, miR-15a/16 and pT3-EF1a-Bmi1 plasmid that lack 3′UTR. Scramble miRNA vector used to serve as a control. Actin is used as a gel loading control. Blots were cropped to enhance the representation (B). Immunofluorescence studies showing accumulation of BMI1, ϒ-H2AX (C), Ub-H2A (D) in MDAMB-231 cells transfected with anti-miR-15a anti-miR-16 or both anti-miR-15a/16 under etoposide- treated conditions. Bar indicates 200 μm. Expression of BMI1, Ub-H2A, γ-H2AX, p-CHK2, RNF8 were checked in MDAMB-231 cells transfected with either scrambled si-RNA or si-RNA specifically against BMI1. Actin is used as a gel loading control. Blots were cropped to enhance the representation (E). Immunofluorescence studies showing the localization of BMI1 and γ-H2AX foci in cells treated with scramble si-RNA (si-scrambled) or si-RNA against BMI1 (si-BMI1). Cells show an increase in γ-H2AX foci upon depletion of BMI1. Bar indicates 200 μm (F).