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. 2013 Oct;33(20):3994–4007. doi: 10.1128/MCB.00623-13

Fig 4.

Fig 4

Repressed KLF3 is required for miR-23a-enhanced globin gene expression. (A) Western blot analysis of KLF3 in K562 cells cotransfected with miR-23a inhibitor and/or KLF3 siRNA after 48 h of hemin induction. (B) qPCR analysis of ε- and γ-globin gene expression in the K562 cells shown in panel A. (C) Western blot analysis of KLF3 in K562 cells transfected with either KLF3 siRNA or pCMV-KLF3. (D and E) qPCR analysis of ε- and γ-globin gene expression and benzidine staining of hemoglobin-containing cells in K562 cells transfected with KLF3 siRNA (D) or pCMV-KLF3 (E) after 48 h of hemin induction. (F) FACS analysis of CD235a/CD71 double-positive cells in K562 cells transfected with KLF3 siRNA or pCMV-KLF3 after 48 h of hemin induction. (G) Schematic of the β-like globin gene locus and the KLF3 binding sites in the HS2 and HS3 regions. (H) ChIP-PCR analysis of KLF3 binding on the CACCC sites in the HS2 and HS3 regions during K562 cell erythroid differentiation. (I) ChIP-qPCR analysis of KLF3 binding on the CACCC sites in the HS2 and HS3 regions during K562 cell erythroid differentiation. Student's t test (two-tailed) was performed to analyze the data from the experiments in triplicate. *, P < 0.05; **, P < 0.01.