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. 2018 Sep 28;10(10):2813–2822. doi: 10.1093/gbe/evy215

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© The Author(s) 2018. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 2. — —GiTim17 is an inner mitosomal membrane protein. (A) GiTim17 was expressed with a C-terminal HA-tag and the protein was detected by western blot of G. intestinalis cellular fractions. The protein was present in the lysate and the high speed pellet fraction, which is enriched for mitosomes. Lys-lysate, Cyt-cytosol, HSP-high speed pellet. (B) Mitosomal localization of GiTim17 was confirmed by immunofluorescence microscopy using GL50803_9296 as the mitosomal marker. (C) STED microscopy of HA-tagged GiTim17 shows its discrete localization on the periphery of the mitosomes, corresponding to the mitosomal membrane. Two images on the left depict details of the displayed cell. (D) Western blot analysis of digitonin-solubilized HSP fraction shows differential distribution of GiTom40 (the outer mitosomal membrane marker) and GiTim17. GiTim17 was found along with GiPam18 and GiTim44, which are associated with the inner mitosomal membrane. S-supernatant, P-pellet.