Fig 3.

KLF3 and SP1 are direct targets of miR-23a and miR-27a, respectively. (A) Computer prediction of the binding of miR-23a and miR-27a on the 3′ UTR of human KLF3 and SP1, respectively. (B) The relative luciferase (LUC) activity of the reporter constructs cotransfected with scrambled control (Neg-control) or miR-23a and miR-27a mimics. Firefly luciferase activity was normalized to the activity of coexpressing renilla luciferase. (C) The relative luciferase activity of the reporter constructs in K562 cells left uninduced or induced with hemin for 48 h. (D) Western blot analysis of KLF3 and SP1 in K562 cells transfected with the scrambled control (NC) or miR-23a and miR-27a mimics and inhibitors. (E) qPCR analysis of the KLF3 and SP1 mRNA levels in K562 cells transfected with the scrambled control or miR-23a and miR-27a mimics. (F and G) The expression of KLF3 and SP1 during erythroid differentiation of hemin-induced K562 cells. (H) Western blot of KLF3 and SP1 in the lentivirus-infected CD34⁺ HPCs after Epo induction for 4, 8, and 11 days. (I) The mRNA level of KLF3 and SP1 during erythroid differentiation of Epo-induced CD34⁺ HPCs. Student's t test (two-tailed) was performed to analyze the data from the experiments in triplicate. *, P < 0.05; **, P < 0.01.