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. 2021 Jun 25;40(31):4955–4966. doi: 10.1038/s41388-021-01904-4

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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 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 — A Percentage of Aldefluor^high (ALDH^high) subpopulations of RUCH-2 cells stably expressing HRAS, KRAS, NRAS or pMSCV (n = 3) measured by flow cytometry. B Representative western blot (n = 2) (left) and corresponding densitometric analyses (right) of ALDH1A1 expression in HRAS-, KRAS-, and NRAS-expressing RUCH-2 cells in comparison to RUCH-2 pMSCV control cells. Protein lysate of murine liver served as positive control. C Mean fold regulation of 84 cancer stem cell-associated genes in HRAS-, KRAS-, and NRAS-expressing RUCH-2 cells compared to RUCH-2 pMSCV cells (n = 2). D MYC qRT-PCR analyses and E anti-SOX2 antibody stainings and percentage of SOX2⁺ nuclei of KRAS and NRAS-expressing RUCH-2 xenotransplants (n = 8 or n = 3 for MYC qRT-PCR or SOX2 stainings, respectively) compared to respective pMSCV control tumors of the same mice (set to 1 for MYC qRT-PCR). Bars in A, D: mean + SEM. Scale bars: 100 µm. Arrows: SOX2⁺ nuclei.