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. 2020 Jun 15;11:3020. doi: 10.1038/s41467-020-16836-y

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© The Author(s) 2020

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 Representative bi-parametric (CD44/CD24) FACS analysis of MCF10A cells expressing the indicated constructs. The CD44^high/CD24^low population is boxed and shown in red, with the relative percentage. See also Supplementary Fig. 10A for a different representation of the results. b The size of the CD44^high/CD24^low population (measured by FACS as in a) in the indicated samples is reported as a percent of the total. Mean ± S.D. of at least two independent experiments is shown (N, Ctr=4, EPN1 = 2, EPN3 = 8, TWIST = 6). P value, each pair Student’s t test two-tailed. c Mammosphere assay on MCF10A cells expressing the indicated constructs. Results are expressed as mean sphere-forming efficiency (SFE) of two independent experiments. P value, each pair Student’s t test two-tailed. d Mammosphere assay on the CD44^high/CD24^low and CD44^low/CD24^high FACS-sorted subpopulations of MCF10A cells expressing the indicated constructs. Results are expressed as mean SFE ± S.D. of two independent experiments, each performed in technical triplicates. P value, each pair Student’s t test two-tailed. e The indicated FACS-sorted populations from MCF10A-EPN3 or -TWIST cells were analyzed by IB as shown and compared to the total unsorted population. GAPDH, loading control. This panel was assembled from samples run on the same gel by splicing out the irrelevant lanes, as shown by the black lines. MW markers are shown on the right. f Representative bi-parametric (CD44/CD24) FACS analysis of the indicated cell lines, mock-silenced (Ctr, top) or silenced for TGFβR1 (bottom). The CD44^high/CD24^low population is boxed and shown in red, with the relative percentage. Results are representative of two independent experiments. g MCF10A cells expressing the indicated constructs were transfected with a non-targeting oligo (scramble), mock-transfected (mock), or silenced for TGFβR1 (TGFβR1 KD) and analyzed by IB. GAPDH, loading control. MW markers are shown on the left. Source data are provided as a Source Data file.