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. 2020 Oct 2;11:4964. doi: 10.1038/s41467-020-18802-0

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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. 4 — a qPCR confirmed the overexpression of miR-1915-3p in CD34⁺-HSPCs after miRNA mimics (miR-1915) were transfected for 24 h compared with the negative control (NC). U6 was set as the normalized control (n = 3 donors). b, c Overexpression of miR-1915-3p promoted the CFU-MKs from the CD34⁺-HSPCs (n = 3 donors, paired-samples t-tests) b and increased the expression of the MK integrins CD41 and CD61 on day 15. (mean ± S.E.M. of n = 5 donors, paired-samples t-tests) (c). Representative FCM plots are shown in Supplementary Fig. 6c. d By megakaryocytic induction on day 20, Cytospin and Wright–Giemsa staining demonstrated more polyploid cells and larger cell dimensions (black arrows) with miR-1915-3p overexpression. Cells from five random views were measured. Statistical analysis and results were conducted on data from three biologically independent experimental replicates Unpaired t-tests with Welch’s correction were used for the statistical analysis. p < 0.0001. (Scale bars: 100 μm) e–g In the K562, Meg-01 and UT-7 cells, overexpression of miR-1915-3p by stable transfection with a plasmid vector carrying primary miR-1915-3p (n = 3 independent experiments) (e) promoted the CD41⁺/CD61⁺ populations (n = 3 independent experiments) (f) when cells were treated with a suboptimal amount of PMA for 3 days. Representative FCM plots are shown in Supplementary Fig. 7a. Cell polyploidization was upregulated with overexpression as well. g The percentage of cells of each ploidy is shown. Representative FCM histograms are shown in Supplementary Fig. 7b (n = 3 independent experiments). h, i Suppression of miR-1915-3p expression with a miRNA sponge (n = 3 independent experiments) h significantly decreased the CD41⁺/CD61⁺ populations in the PMA-treated K562, UT-7 and Meg-01 cells i Representative FCM plots are shown in Supplementary Fig. 7d (n = 3 independent experiments). All data are shown as the mean ± S.D from two-tailed unpaired-samples t-tests. P-value unless otherwise specified. Source data are provided as a Source Data file *P < 0.05, **P < 0.01, ***P < 0.001.