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. 2021 May 24;11:10791. doi: 10.1038/s41598-021-89931-9

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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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The effect of inhibition of MDR1 and ABCG2 on DOX-induced apoptosis. The cells were treated with 1 µM of MDR1 inhibitor valspodar or 1 µM of ABCG2 inhibitor ko143, and doxorubicin (DOX) (200 nM for Huh7 and 100 nM for PLC/PRF/5 cells, respectively) for 24 h. Alternatively, the cells were treated with 20 nM siRNA to MDR1 or ABCG2 for 6 h, followed by a further 42 h culture in the fresh culture medium. Then the Huh7 cells or PLC/PRF/5 cells were treated with 200 nM or 100 nM of DOX for 24 h. The total percentage of apoptotic cells as defined by the combination of 7-AAD⁻/Annexin V⁺ and 7-AAD⁺/Annexin V⁺ cells are shown for (a) bulk Huh7 cells, (b) bulk PLC/PRF/5 cells, (c) EpCAM⁺–CD133⁺ Hun7 cells as well as (d) EpCAM⁺-CD133⁺ PLC/PRF/5 cells. Data shown are mean ± SD, n = 3. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; compared with DOX-only treatment.