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. 2017 Nov 27;8:1786. doi: 10.1038/s41467-017-01690-2

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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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Fig. 3 — Effects of a pharmaceutical and a genetic inhibition of HDAC6 on terminal human MK differentiation. a ACY1215 and Tubastatin A decreases PPF. b Confocal analysis of untreated and treated MKs. Proplatelets derived from treated MKs display shorter extensions compared to untreated MKs. Bar represent 10 µm. c sh1 and sh2 leads to a specific inhibition of PPF. Results are representative of three independent experiments (n = 3). Unpaired Student’s t test:**p < 0.001; ***p < 0.0001. d Ultrastructure analysis of shC and shHDAC6 MKs by transmission electron microscopy. (i, ii) Two MKs with a normal ultrastructure in the sh control with an enlargement, a regular development and distribution of demarcation membrane system (DMS), α-granules (a) and multivesicular bodies (MVB). We can also observe normal proplatelet (PPT) produced in vitro. (iii, iv) Two examples of abnormal ultrastructure of shHDAC6-treated MKs. Enlargement on both shHDAC6 MKs reveal either (iii) an abnormal presence of cytoplasmic vacuoles containing numerous microvesicles looking to large MVB and giant α-granules, or (iv) mature MKs displaying an heterogeneous granules size content. e Confocal analysis of CD63 and vWF in MKs transduced with shC or with shHDAC6. Bar represent 30 µm. Error bars are SEM