Figure 3

Erythroid-specific downregulation of target gene expression in the erythroid cells differentiated from CD34⁺ HSPCs transduced with Ery-Lin-shRNA LVs. (A) Schematic outline of the experiment. (B) Representative immunoblots that illustrate the knockdown efficiencies of shBCL11A and shZBTB7A in the erythroid cells (left) and the densitometric data of the immunoblots (right). ZG represents ZsGreen expression; ‘+’ indicates ZsGreen⁺ cells and ‘–’ indicates ZsGreen^- cells. Full blots are presented in Supplemental Fig. S9 and S10. (C) Intracellular HbF analysis by flow cytometry to determine the percentages of HbF⁺ cells in the erythroid cells differentiated from the shRNA transduced human CD34⁺ HSPCs  (left) and the graphical representation of the data (right). (D) HPLC analysis of the percentages of Gγ+Aγ chains in the shRNA transduced cells after differentiation (left) and the graphical representation of the data (right). H23B and H234B represent H23B-Ery-Lin-shRNA and H234B-Ery-Lin-shRNA LVs, respectively. All data are mean ± SD (n = 2).