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. 2021 Dec 15;12:7300. doi: 10.1038/s41467-021-27308-2

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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. 8 — a Clinical and pathological characteristic of the 49 breast cancer cases included in the analysis. b A schematic of tissue collection procedure for the correlation of TMEM doorway score with the proportion of cancer cells expressing stem cell markers. c TMEM correlation with stem cell marker CD44+/24− in 49 invasive ductal carcinomas. Cells were collected by FNA and analyzed by flow cytometry for % cells expressing CD44+/CD24−. TMEM score from the same tumor tissue was correlated with the % CD44+/24− cells by Pearson’s correlation. d Correlation of TMEM with the percentage of breast cancer cells expressing stem cell markers CD44+/24−, ALDH1, and CD133 in 49 invasive ductal carcinomas from patients. Cells were collected by FNA and analyzed for CD133 and ALDH1 by in situ hybridization. CD44+/24− data are plotted from c. TMEM score from the same tumor tissue was correlated with the percentage of cancer cells expressing stem cell markers using Pearson’s correlation. e Correlation of TMEM with the percentage of breast cancer cells expressing stem cell markers ALDH1 and CD133 in 48 invasive ductal carcinomas from patients. Cells were collected by FNA and analyzed for CD133 and ALDH1 by qRT-PCR. TMEM score from the same tumor tissue was correlated with the percentage of cancer cells expressing stem cell markers using Pearson’s correlation. f Sox9 mean fluorescence intensity in FFPE of the above human tissues (n = 13) were correlated with the TMEM score for each human tissue. Pearson’s correlation r = 0.86, p = 0.0002, data normalized to the min and max Sox9 mean fluorescence intensity values. g A composite image of TMEM staining (pink: Mena+ tumor cells, brown: CD68+ macrophages, blue: CD31+ endothelial cells) and Sox9^Hi stem cells (green), showing enrichment of Sox9^Hi cells close to TMEM areas. Two TMEM doorway ROIs are highlighted with black circles. h Quantification of % Sox9^Hi cells in TMEM-rich and TMEM-deficient areas. n = 3 human tissues, data plotted as mean ± SD, paired two-tailed Student’s t-test, *p = 0.0336.