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. 2020 Jun 12;11:2807. doi: 10.1038/s41467-020-16179-8

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© The Author(s) 2020

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. 1 — a Relative NSD1 mRNA expression (1/dCt) in peripheral blood CD34⁺ cells transduced with pLKO.1 expressing control shRNA (Ctrl) or NSD1 shRNA (#372) harvested from the first and second plating in growth-factor-containing MC (H4434). Bars represent average relative expression normalized to (n = 5 per group). b Numbers of colonies formed by 4 × 10⁴ peripheral CD34⁺ cells transduced with pLKO.1 expressing control shRNA (Ctrl) or NSD1 shRNA in the first plate (n = 5) and upon replating (n = 2) in growth-factor-containing MC (H4434). c Representative images of colonies formed in MC (H4434) by 4 × 10⁴ peripheral CD34⁺ cells transduced with pLKO.1 expressing control or NSD1 shRNA. d Flow cytometric analysis of cells harvested from the first and second plating in MC (H4434) revealed accumulation of CD71^high and glycoprotein A (GPA)⁻ cells upon replating. The plots represent one out of three independent experiments. e Representative images of Wright Giemsa-stained cytospin preparations from control shRNA (Ctrl) or NSD1 shRNA-expressing CD34⁺ cells harvested from the MC (H4434) cultures after the first and second plating, illustrating the overall predominance of cells with erythroblast morphology upon replating (one out of three experiments) (×1000, the size bar = 10 μM). Values are presented as individual points, bar graphs represent the mean value of biological replicates, error bars as standard error of the mean. Statistical significances in a, b was tested with paired two-tailed t-test.