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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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Effect of the inhibition of MDR1 and ABCG2 on the capability of doxorubicin (DOX) in the elimination of cancer stem cells in vitro. Huh7 or PLC/PRF/5 cells were treated with either doxorubicin (DOX) alone or treated with inhibitors or siRNA to MDR1 or ABCG2 first, followed by DOX treatment as described in “Materials and methods” section. Cells were then plated in ultra-low attachment plates at a density of 10, 20, or 50 cells/well and incubated for 7 days under the condition of in vitro limiting dilution assay to assess the self-renewal capacity of treated cells. The frequency of tumorsphere-forming cells for Huh7 (a) or PLC/PRF/5 cells (b) under indicated treatment is shown. Data presented are mean ± SD, n = 3. *p < 0.05; ***p < 0.001.