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. 2019 Oct 21;1(2):161–167. doi: 10.1097/BS9.0000000000000031

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Copyright © 2019 The Authors. Published by Wolters Kluwer Health Inc., on behalf of the Chinese Association for Blood Sciences.

This is an-open access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. http://creativecommons.org/licenses/by-nc-nd/4.0

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Identification and functional verification of lncRNA PCED1B-AS1. (A) Dynamic expression of lncRNA PCED1B-AS1 during erythroid differentiation in Blueprint database. (B) Relative expression of PCED1B-AS1 during late stage of erythroid differentiation of CD34+ cells. The horizontal axis represents the days after EPO induction. (C) The confirmation of PCED1B-AS1 overexpression in K562 cells by RT-PCR. D3 represents the cells induced with hemin for 3 days. (D) The analysis for globin gene expression changes before and after induction of PCED1B-AS1-overexpressed K562 cells compared with that in control cells. HBE, HBG and HBB are β-like genes that respectively represent ε (embryonic)-, γ (fetal)- and β (adult)-globin and are specifically expressed at different developmental stages during erythroid differentiation. (E) The expression changes of CD235a and CD71 was detected by flow cytometry before and after induction. The percentage in the upper right of each figure represents the proportion of CD235a⁺-CD71⁺ double positive cells. (F) Percentage of CD235a+-CD71+ double positive cells among the K562 cells before and after induction. Data are the means ± SD of three experimental replicates. N.S. = no significance between samples. (G,H) Benzidine-stained positive rate changes of K562 cells before and after induction with hemin. All relative mRNA expression was normalized to GAPDH. Statistical results were analyzed by student t-test and Kruskal−Wallis test, ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ^nsP > .05, ns: no significance. D0: Day0, D3: Day3. Ctr: Control, OE: overexpression of PCED1B-AS1.