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. 2018 Feb 22;8:3495. doi: 10.1038/s41598-018-21409-7

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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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Characterization of bone marrow stromal cells (BMSCs). CD45⁻ BMSCs (gate R3) of Lgals3^+/+ (A) and Lgals3^−/− mice (B). (C) The relative expression of genes involved with hematopoietic events was compared between CD45⁻ BMSCs of Lgals3^+/+ and Lgals3^−/− mice (Lgals3^−/− per Lgals3^+/+). (D) Total bone marrow cells were cultured in the methylcellulose system containing B cell-enriched growth factors: colonies (maximum 50 cells) and clusters (minimum 50 cells). (E) Coculture systems using different strategies to quantify B220⁺CD19⁺ B cells: BM Lgals3^+/+ (culture of total bone marrow cells of Lgals3^+/+); BM Lgals3^+/+ in Lgals3^−/− (bone marrow cells of Lgals3^+/+ mice cultured in BMSCs of Lgals3^−/−); BM Lgals3^−/− in Lgals3^+/+ (bone marrow cells of Lgals3^−/− cultured in Lgals3^+/+ BMSCs); BM Lgals3^−/− (culture of total bone marrow cells of Lgals3^+/+). (F) Number of B220^lowIL-7R⁺ cells after 1 week using similar co-culture strategy. (G–I) Histograms represent the expression of Delta-like 1 (DLL1), Delta-like 4 (DLL4) and Jagged-1 (JAG1) by CD45⁻ BMSCs (gated in R3, A and B). n = 5 mice per group. (*) Indicates p < 0.05. Dot plot graphs are representative of each mice group.