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. 2017 Jun 8;8:15710. doi: 10.1038/ncomms15710

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Copyright © 2017, The Author(s)

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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(a) WB performed on total extract of extracellular tachyzoites, wild type (ΔKU80) or expressing the endogenously tagged TgUNC (UNC-3Ty). Actin (ACT) was used as loading control. (b) UNC-3Ty was detected in the cytosol of intracellular tachyzoites co-stained with the peripheral marker GAP45. Scale bar, 2 μm. (c) The regulation of the Tet-inducible cell line of TgUNC (MycUNC-iKD) was assessed by WB on total extract of extracellular tachyzoites. ACT was used as loading control. (d) TgUNC depletion has a severe impact on parasite survival as shown by plaque assay treated for 7 days±ATc. (e) All classes of myosin heavy chains are destabilized on TgUNC depletion. All the myosin motors, except TgMyoA and TgMyoD, have been endogenously tagged with 3xTy in the MycUNC-iKD strain and their expression level was followed by WB on total extracts of extracellular tachyzoites treated or not with ATc for 24 or 48 h. Expression of TgMyoD was followed using α-MLC2 antibodies. GAP40 or ACT were used as loading controls. MycUNC-iKD was tightly regulated in all samples. Stars indicate myosin subproducts. Uncropped gels are presented in Supplementary Fig. 11.