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. 2021 Mar 11;11:5752. doi: 10.1038/s41598-021-85056-1

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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 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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MAGI1 impairment induces tumorigenic phenotypes in epithelial cells. (A) MTT assay (OD 560 nm) representing 2D cell growth of MCF7shMAGI1 as compared to MCF7shLuc cells. Bars represent mean ± Standard Deviation (SD; n = 10 wells as replicates) of a representative experiment (out of 3). Unpaired two-tailed Student's t-test; **p < 0.01; ***p < 0.001. (B) Cell cycle phases of MCF7shLuc and MCF7shMAGI1 cells assessed after BrdU incorporation and analyzed by flow cytometry, showing an increased proportion of cells in S phase at the expense of G0/G1 after MAGI1 invalidation. The pie chart also showed no major difference in the percentage of apoptotic cells in SubG1 phase (around 2% in shLuc and 1% in shMAGI1 cells). (C) Left: quantification of colony numbers of MCF7shMAGI1 cells grown in anchorage independent conditions (soft agar assay) and represented as fold increase compared to MCF7shLuc cells. Data are presented as the means ± SD (n = 3). Unpaired two-tailed Student's t-test; ***p < 0.001. Right: western blot analysis of whole protein extracts issued from the cells used in the soft agar assays and showing the relative amounts of MAGI1 in the different cell lines; Tubulin was used as a loading control. Uncropped blots can be found in the Supplementary Information. (D) Top: quantification of mammospheres represented as total area issued from MCF7shMAGI1 cells and shown as fold increase compared to MCF7shLuc cells, starting from 200 cells. Data are presented as the means ± SD (n = 4). Unpaired two-tailed Student's t-test; ***p < 0.001. Bottom: images of mammosphere formation assay showing a representative experiment.