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. 2023 Feb 23;23:33. doi: 10.1186/s12935-023-02874-1

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© The Author(s) 2023

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — KLF9 restricts breast epithelial cell survival and proliferation. Effects of CORT treatment and KLF9 knockdown (blue) or overexpression (yellow) on A, C, E cell survival and B, D, F viability of breast epithelial cell lines as evaluated using assays based on colony formation and resazurin reduction, respectively. The cells were treated with vehicle (100% ethanol) or CORT (100 nM) for 14 d and 96 h for the colony formation and cell viability assay, respectively. In MCF10A, CORT promoted A survival and B proliferation in both knockdown [2-way ANOVA; Colony formation: treatment: P < 0.0001; knockdown: P = 0.2060|Cell viability: treatment: P < 0.0001; Knockdown: P = 0.0004] and overexpression lines [2-way ANOVA; Colony formation: treatment: P < 0.0001; overexpression: P = 0.0005|Cell viability: treatment: P = 0.1045; overexpression: P < 0.0001. In the luminal MCF7 line, CORT treatment diminished C survival and D proliferation in both knockdown [2-way ANOVA; Colony formation: treatment, knockdown: P < 0.0001|Cell viability: treatment: P < 0.0001; knockdown: P = 0.0355] and overexpression lines [2-way ANOVA; colony formation: treatment: P < 0.0001; overexpression: P = 0.0269|Cell viability: treatment: P < 0.0001; overexpression: P < 0.0001]. Finally, in MDA-MB-231 cells, CORT treatment enhanced E colony formation and F proliferation in both KLF9 knockdown [2-way ANOVA; colony formation: treatment, P = 0.1026; knockdown: P = 0.0002|Cell viability: treatment: P = 0.0006; Knockdown: P = 0.0057] and overexpression lines [2-way ANOVA; Colony formation: treatment: P = 0.0986; overexpression: P < 0.0001|Cell viability: treatment, overexpression: P < 0.0001]. Bars represent mean ± SEM with statistically significant differences determined through two-way ANOVA for main effects of treatment or overexpression and Student’s t-test for individual effects of hormone treatment (^*) and knockdown or overexpression (^#). All experiments were performed with N ≥ 4 replicates