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. 2024 Mar 13;10:29. doi: 10.1038/s41421-024-00655-2

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© The Author(s) 2024

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 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/.

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graphic file with name 41421_2024_655_Figg_HTML.jpg — Figure 4 Modulation of calcium-binding protein 39-like (CAB39L) and Sestrin-1 (SESN1) underlies metformin anticancer activities. (a–c) Colony-forming assay. Histograms showing average colony counts of SUM159PT (a), BT-474 (b) and MCF-7 (c) cells expressing either a control vector or a CAB39L-expressing vector or CAB39L- or SESN1-targeting shRNAs and treated with vehicle or metformin (0.5 mM) before seeding at clonal density. Statistics (t-test): P < 0.05. (d) Wound healing assay. Percentage of wound closure (over vehicle) of SUM159PT cells transfected and treated as from panel (a–c), for 48 h. Statistics (t-test): P < 0.05. (e, f) Invasion assay. BT-474 (e) and MCF-7 (f) cells were treated with vehicle or metformin (0.5 mM) for 24 h and the number of migrated cells was scored. (g, h) Colony-forming assay (g) and invasion assay (h). Histograms showing average colony counts (g) or the number of migrated cells (h) of SUM159PT cells transfected either with a control vector or CAB39L- or SESN1-targeting shRNAs and treated with vehicle or metformin (0.5 mM). Histograms indicating the average ± s.e. of triplicate experiments. Statistics (t-test): P < 0.05. *Significant versus vehicle, ºSignificant versus untransfected, metformin-treated ones.