Modulation of red cell maturation by multiple types of lncRNAs. (A) Relative expression of the early erythroid differentiation marker TER119 in erythroid progenitor-enriched fetal liver cells transduced with retroviral vectors encoding control or lncRNA-targeting shRNAs and induced to differentiate in culture. Expression levels were determined by qPCR, normalized to those of 18S rRNA, and are shown as percentage of the levels in the control shRNA experiment (dotted gray line). Data are mean ± SEM (n = 2). (B) Relative cell size of cells treated as in A. Average cell sizes were determined by flow cytometry (Methods) and are shown as percentage of the values for the control shRNA experiment (dotted gray line). Data are mean ± SEM (n = 2). (C) Relative enucleation efficiency of cells treated as in A. Enucleation efficiency was determined by flow cytometry (Methods) and is shown as percentage of the values for the control shRNA experiment (dotted gray line). Data are shown as mean ± SEM (n = 2). (D) (Top left) SPRYD7 (light gray) is anticorrelated in expression with its neighbor shlncRNA-EC6 (dark gray) during erythropoiesis. (Top right) depletion of shlncRNA-EC6 with separate shRNAs in ex vivo–differentiated TER119⁺ erythroblasts results in reproducible up-regulation of SPRYD7 relative to scramble shRNA control (data are mean ± SEM; n = 3). (Bottom left) KIF2A (light gray) is coordinated in expression with neighboring elncRNA-EC3 (dark gray) during erythroid differentiation. (Bottom right) inhibiting elncRNA-EC3 with separate shRNAs leads to reduced expression of KIF2A relative to scramble shRNA control (data are mean ± SEM; n = 3).