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. 2021 Nov 2;11:21425. doi: 10.1038/s41598-021-00954-8

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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(A) Results of real-time PCR analysis of MDA-MB-231 cells treated with all formulations (0.259 μg/mL of B2D, 2D, BD, Dox, BC and Conf and 2.59 μg/mL of PB) as the heat map of expression level of genes related to un-treated control group (gene expresion = 1). The yellow and light-yellow colors are the signs of lack of gene expression or downregulation of it and red color shows upregulation of gene expression related to the control (genes: Bcl-2, Bax, Caspase-3, Caspase-6, Caspase-7, Caspase-8, Caspase-9, Caspase-10, Caspase-12 and GAPDH as the internal control gene); (B) Diagram of proteins expression changes related to the control group (protein expression = 1) obtained from western blotting (the MDA-MB-231 cells were treated by B2D with concentration of 0.259 μg/mL). Proteins: Bcl-2, Bax, pro-Caspase-9, Cleaved-Caspase-9, pro-Caspase-3, Cleaved-Caspase-3, pro-Caspase-7, Cleaved-Caspase-7, p27 and p53, and GAPDH as internal control, n = 2; (C) Images of western blotting of the MDA-MB-231 cells treated with B2D (0.259 μg/mL). Un-treated cells were considered as the control group. (Abbreviations: PB: blank βCD-g-PMA-co-PLGA micelles, B2D: co-drug loaded βCD-g-PMA-co-PLGA micelles, BD: Dox loaded βCD-g-PMA-co-PLGA micelles, BC: Conferone loaded βCD-g-PMA-co-PLGA micelles, Dox: Free Doxorubicin, Conf: Free Conferone, 2D: Free Doxorubicin-Conferone).