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. 2011 Jul 28;6(7):e22907. doi: 10.1371/journal.pone.0022907

Figure 2. Overexpression of Eos reduces γ-globin gene expression in μ'LCRAγψβδβ/GM979 cells.

Figure 2

(A) Structural diagram of the integrated construct μ'LCRAγψβδβ, in which a 3.1-kb μ'LCR cassette is linked to a 29-kb fragment from the human Aγ- to β-globin gene. (B) Eos protein level was analyzed by Western blot in μ'LCRAγψβδβ/GM979 cells that were untreated and transfected with pcDNA3.1 or pcDNA-Eos. (C) The expression levels of human γ-, β-, and murine α-globin genes were analyzed by Northern blot in stable μ'LCRAγψβδβ/GM979 transformants. Each globin mRNA level was normalized to murine β-actin mRNA. M, C, and E represent μ'LCRAγψβδβ/GM979 cells that were untransfected, transfected with pcDNA3.1 (control), and transfected with pcDNA3.1-Eos, respectively. (D) Human γ-, β-, and murine α-globin mRNA levels were measured by quantitative real-time PCR. Each PCR analysis was performed in triplicate, and expression levels were normalized to murine β-actin mRNA. Error bars represent standard deviation. *P<0.05.