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. 2018 Nov 21;8:17174. doi: 10.1038/s41598-018-33873-2

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© The Author(s) 2018

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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The 100 kDa NCL glycoform is expressed on the surface of pre-B ALL cells. Analysis of the indicated pre-B ALLs using NCL antibody. (a) Western blotting on cell surface (biotinylated) and intracellular (non-biotinylated) fractions. Note: Different WB membranes of the two fractions; the WB of the intracellular fraction was exposed shorter than that of the cell surface fraction. (b–e) Flow cytometry. (b) Representative analysis of cell surface NCL in different pre-B ALLs. (c) Summary graph of n = 2–5 independent experiments with the indicated pre-B ALLs. (d) NCL expression in gate P1 on primary fresh pre-B ALL PB sample containing >95% blasts. For FACS, cells were fixed without permeabilization. (e) Analysis for cell surface NCL and 9-O-Ac-Sia on normal pre-B cells. Percentage of cells in five different normal bone marrow samples, processed as described in Materials and Methods, that were positive for cell surface NCL and 9-O-Ac-Sia in the CD19+, CD10+ gate (gating strategy, see Supplementary Fig. S3). ICN13, positive control run in the same experiment.