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. 2018 Apr 20;8:105. doi: 10.3389/fonc.2018.00105

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Copyright © 2018 Di Ianni, Baldoni, Del Papa, Aureli, Dorillo, De Falco, Albi, Varasano, Di Tommaso, Giancola, Accorsi, Rotta, Rompietti, Silva Barcelos, Campese, Di Bartolomeo, Screpanti, Rosati, Falzetti and Sportoletti.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of NOTCH1-TM expression in immunoselected CD34+ bone marrow (BM) cells. (A) Immunomagnetic cell sorting purification and phenotypic characterization of CD34+ cells from a representative chronic lymphocytic leukemia (CLL) BM used for NOTCH1 protein expression analysis. (B) (i and ii) Western blot analysis. Vertical lines indicate realignment of the same blot imaging. Protein loading was assessed by reprobing the blots with an anti-GAPDH antibody. The density of the bands corresponding to NOTCH1-TM was evaluated by densitometric analysis. Densitometry units (U) were calculated relative to GAPDH. Results revealed that CD34+ samples from CLL patients always expressed the NOTCH1-TM protein. (C) Schematic representation of clonal evolution during the development of CLL starting from hematopoietic stem cells (HSCs). NOTCH1 is found to be active at early stage of hematopoiesis (blue arrow) together with NOTCH1 mutations (lightning arrow) to which activated signaling might contribute. NOTCH1 activation persists and the mutated clone expands as the cell commit to mature CD19+CD5+CLL.