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. 2014 Apr 4;28(9):1811–1818. doi: 10.1038/leu.2014.100

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Copyright © 2014 Macmillan Publishers Limited

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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Immunostaining of bone marrow biopsies with the anti-mutated CALR antibody. Representative sections from CALR-unmutated ET/PMF patients (JAK2V617F mutated, MPLW515L mutated and triple-negative mutation) and from CALR-mutated patients (CALRdel52, CALRins5 and CALRindel) are shown in panel a and panel b, respectively. Sections were stained according to the procedure described in the Materials and Methods section using a 1:1000 dilution of the anti-mutated CALR antibody. Pictures were taken with a LEICA DM LS2 microscope using a N-Plan × 40/0.65 objective. The number of immunostained megakaryocytes over total number of morphologically recognizable megakaryocytes was calculated by counting at least 100 megakaryocytes/slide from 10 patients with ET and 10 patients with PMF (n=2 for the CALR indel), and expressed as percent of total megakaryocytes (c); as there was no significant difference between the two groups, all individual data were pooled.