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. 2017 Sep 26;7:12297. doi: 10.1038/s41598-017-12069-0

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© The Author(s) 2017

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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Identification of mitochondria in mature RBCs of RTT patients by IF. Upper panel: Immunofluorescence microscopy analysis of the RBCs isolated from the RTT patients carrying the R255X MeCP2 mutation (n = 3) (left panel) and from healthy individuals (n = 3) (right panel). RBCs were allowed to adhere to glasses by cytospinning and were probed with an anti COX-IV antibody. RTT RBCs displayed a COX IV⁺ dotted pattern that was undetectable in healthy RBCs. Clear field acquisition highlighted a normal bi-concave shape of the RBCs. The experiment was carried out in triplicate by using the same blood specimen. Lower panel: the percentage of RBCs displaying at least 1 mitochondria was calculated in 10 different fields (left panel); the average number of mitochondria per RBC was calculated by counting the number of dots for each RBC in the different fields (right panel). The results are the means ± S.E.; **significantly different from control (**p < 0.005, unpaired τ Student’s test).