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. 2020 Jul 13;11(7):530. doi: 10.1038/s41419-020-2719-2

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© The Author(s) 2020

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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Fig. 4 — CCK-8, colony formation, DNA replication activity and apoptosis assays for MDA-MB-231/DR and MDA-MB-231/VR cells cultured with control, 5 mM aspirin (Asp), 5 nM docetaxel (Doc), 5 μM vinorelbine (Vin), aspirin plus docetaxel (A + D), and aspirin plus vinorelbine (A + V). The rest of the assays were performed as and the groups were similar to those described above. a CCK-8 assay for MDA-MB-231/DR. Control vs A + D (P < 0.001); Asp vs A + D (P = 0.004); Doc+ A + D (P = 0.002). b CCK-8 assay for MDA-MB-231/VR. Control vs A + D (P = 0.001); Asp vs A + D (P = 0.036); Doc+ A + D (P = 0.002). b CCK-8 assay for MDA-MB-231/VR. Control vs A + D (P < 0.001); Asp vs A + D (P = 0.004); Doc+ A + D (P = 0.002). c Colony formation assay for MDA-MB-231/DR. Control vs A + D (P = 0.001); Asp vs A + D (P = 0.002); Doc+ A + D (P < 0.001). d Colony formation assay for MDA-MB-231/VR. Control vs A + D (P < 0.001); Asp vs A + D (P = 0.005); Doc+ A + D (P = 0.002). e DNA replication activity assay for MDA-MB-231/DR. DNA replication activity is a critical index for cell growth. BrdU, a synthetic thymidine analogue that binds to replicating DNA, was used to examine the rate of DNA replication. A red color indicates cells in the DNA replication phase, and a blue color indicates the cell nucleus. Scale bar: 20 µM. Control vs A + D (P < 0.001); Asp vs A + D (P = 0.002); Doc+ A + D (P < 0.001). f DNA replication activity assay for MDA-MB-231/VR. Control vs A + D (P < 0.001); Asp vs A + D (P = 0.001); Doc+ A + D (P < 0.001). g Apoptosis assay for MDA-MB-231/DR. Control vs A + D (P < 0.001); Asp vs A + D (P = 0.002); Doc+ A + D (P < 0.001). h Apoptosis assay for MDA-MB-231/VR. Control vs A + D (P < 0.001); Asp vs A + D (P < 0.001); Doc+ A + D (P < 0.001). i Western blot for YAP, β-catenin, cyclin D1, c-myc, Bcl-2 and Bax in MDA-MB-231/DR and MDA-MB-231/VR cells cultured with control, 5 mM aspirin (Asp), 5 nM docetaxel (Doc), 5 μM vinorelbine (Vin), aspirin plus docetaxel (A + D), or aspirin plus vinorelbine (A + V) for 48 h. The data are presented as the mean ± SD of experiments performed in triplicate. Statistical significance was determined using Student’s t test analysis.