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. 2021 Mar 17;12(4):286. doi: 10.1038/s41419-021-03555-5

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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. 1 — a Representative immunohistochemical staining of p97 in human breast, colon, liver, and pancreatic cancers and adjacent noncancerous tissues. Bar: 20 μm. b Representative immunohistochemical staining of high-level p97 (red) co-stained with CD44 (brown, left) and CD24 (brown, right) in two consecutive serial sections of human breast cancer tissues. c Representative immunohistochemical staining of low-level p97 (red) co-stained with CD44 (brown, left) and CD24 (brown, right) in two consecutive serial sections of human breast cancer tissues. Bar: upper 200 μm, lower 50 μm. Note that p97 primarily localized to CD44⁺/CD24⁻ cells but not other cells. d Percentages of p97 expression in CD44⁺/CD24⁻ cells and non-CD44⁺/CD24⁻ (CD44⁻/CD24⁻, CD44⁻/CD24⁺, and CD44⁺/CD24⁺) cells in breast cancer tissues (n = 75). e Immunohistochemical staining of SOX2 and p97 in the consecutive serial sections of breast cancer tissues. SOX2 and p97 expressions were classified as low and high. Bar: upper 200 μm, lower 50 μm. f Correlation analyses of SOX2 and p97 expression in breast cancer tissues (n = 75). g Immunoblotting of p97 in lysates of MCF10A, MCF-7, and MDA-MB-231 cells. GAPDH was used as a loading control. h Left: flow cytometry analysis of CD44 and CD24 in MCF10A, MCF-7, and MDA-MB-231 cells. Right: the CD44⁺/CD24⁻ percentages. i qPCR analysis of P97 mRNA in CD44⁺/CD24⁻, non-CD44⁺/CD24⁻, ALDH⁺, ALDH⁻, PKH26⁺, and PKH26⁻ populations. j, k Immunoblotting analysis of p97, OCT4, and SOX2 in CD44⁺/CD24⁻, CD44⁻/CD24⁻, ALDH⁺, and ALDH⁻ populations isolated from MCF-7 and MDA-MB-231 cells. GAPDH was used as a loading control. Data were shown as mean + SD. *P < 0.05 and ***P < 0.001.