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. 2020 Jul 21;10:12109. doi: 10.1038/s41598-020-68694-9

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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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Experimental workflow to induce multinuclearity. The three conditions start from the same amoeba inoculum. (a) Non-agitated Adherent conditions. Amoebae are incubated without agitation throughout the experiment. (b) Agitated Non-adherent conditions. Amoebae are first incubated with agitation for 24 h and then without agitation for 24 h. (c) Agitated Adherent conditions. Amoeba cells are allowed to adhere for 3 h prior to be incubated with agitation for 24 h and then without agitation for 24 h.